Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
This report is prepared according to the 2015 fiscal year of the Royal Thai Government, from 1 October 2014 – 30 September 2015.
2 Annual Report 2015 3 National Center for Genetic Engineering and Biotechnology (BIOTEC) International Collaboration
Annual Report 2015 CONTENTS National Center for Genetic Engineering and Biotechnology (BIOTEC) Message from the BIOTEC Executive Director 5 Facts and Figures 7 Research and Development 11 ISBN 978-616-12-0431-0 Technology Transfer 35 First Edition January 2016 February 2016 Human Resources Development 41 Number of copies printed 500 Public Awareness 47 International Collaboration 51 Copyright ©2016 by Impact of BIOTEC’s Output 57 National Center for Genetic Engineering and Biotechnology (BIOTEC) Appendices 63 National Science and Technology Development Agency (NSTDA) List of Publications 64 List of Intellectual Properties 80 Honors and Awards 86 Executives and Management Team 88
Annual Report 2015 National Center for Genetic Engineering and Biotechnology/ National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency (NSTDA). -- Pathum Thani : National Center for Genetic Engineering and Biotechnology, 2016. 90 p. : ill. ISBN : 978-616-12-0431-0 F
1. Biotechnology 2. Genetic Engineering I. National Center for Genetic Engineering and Biotechnology II. National Science and Technology Development Agency III. Title
TP248.2 660.6
Published by National Center for Genetic Engineering and Biotechnology (BIOTEC) National Science and Technology Development Agency (NSTDA) Ministry of Science and Technology 113 Thailand Science Park Phahonyothin Road Khlong Nueng, Khlong Luang Pathumthani 12120 THAILAND Tel: +66 (0) 2564 6700 Fax: +66 (0) 2564 6701-5 Website: http://www.biotec.or.th International Collaboration 3
CONTENTS
Message from the BIOTEC Executive Director 5 Facts and Figures 7 Research and Development 11 Technology Transfer 35 Human Resources Development 41 Public Awareness 47 International Collaboration 51 Impact of BIOTEC’s Output 57 Appendices 63 List of Publications 64 List of Intellectual Properties 80 Honors and Awards 86 Executives and Management Team 88
F 4 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Message from the BIOTEC Executive Director 5
MESSAGE FROM THE BIOTEC EXECUTIVE DIRECTOR
BIOTEC has always strived to find a balance between Human capital is the most important asset of any excellence and relevance. In Thailand, science is not organization. This year, a number of our staff have intrinsically perceived as relevant, so it is always brought pride to BIOTEC. Just a few examples gratifying when we can see our research results make illustrating the strength and diversity of our human their ways to actual users. This year, we launched a resources; Dr. Bunpote Siridechadilok was named the collaborative project with the Department of 2014 Young Technologist by the Foundation for the Agricultural Extension to transfer the effective Promotion of Science and Technology under the Beauveria bassiana strain and the cultivation Patronage of His Majesty the King for the development technology to the field stations of the Department, of a quick and efficient method to construct mutations as well as the training of farmers to effectively produce of dengue virus which will enable high throughput the fungal spores from the inoculum distributed by screening of drug targets as well as understanding the Department. The effective strain and cultivation gene functions. Dr. Noppol Kobmoo won the Marie process were the fruit of our research program to find Skłodowska-Curie Individual Fellowship to work with useful applications for the wealth of the country’s Prof. Titiana Giraud at the University of Paris-Sud, to biodiversity. B. bassiana has proven to be a potent use population genomics approach to explain the host biopesticide against insect pest larvae. specificity in insect fungi. In addition, Dr. Chalermpol Kirdmanee and Ms. Phongphan Ek-arwut were BIOTEC is currently undertaking two large-scale recognized for their work in the rural areas. Dr. projects under NSTDA Giga Impact Initiative, a new Chalermpol received the Siam Cement Group Chair Fprogram set up in 2013 providing funds to projects Professor Scholarship for the work on land with high potential to generate a significant impact rehabilitation in northeastern Thailand. The National to the economy. One project aims to use genomic Office of Buddhism honored Ms. Phongphan for her technologies in a sugarcane breeding program to support in education of monastic schools in the rural increase yield and sugar level. The other project aims area. Indeed, we never forget to be relevant. to develop a platform for large-scale production of industrial enzymes, concentrating on three This year, the Ministry of Science and Technology has recombinant enzymes for three distinct industries, been regrouped with the economic ministries, a good namely animal feed, textile and pulp and paper. In indication that our policy makers see the potential of addition to the R&D component, both projects are science and technology. It also comes with engaging industrial partners to expedite the transfer responsibilities to deliver innovations to support our of technology to the private sector once completed. local industry. At BIOTEC, we are looking forward to The sugarcane project is co-funded by and co- making this contribution in the years to come. investigated with an industrial partner, Mitr Phol Group, a key player in sugarcane and sugar business in Thailand and the Asia Pacific; whereas the enzyme team is now in discussion with various enzyme users for field validation and commercial enzyme production in Thailand.
Kanyawim Kirtikara, PhD. Executive Director, BIOTEC 6 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Facts and Figures 7
FFACTS AND FIGURES
BIOTEC was first set up under the Ministry for As a premier research institute in Thailand and Asia, Science, Technology and Energy on 20 September BIOTEC operates research units located at Thailand 1983. After the establishment of the National Science Science Park and specialized laboratories hosted and Technology Development Agency (NSTDA) on 30 by various universities, covering a wide spectrum December 1991, BIOTEC became one of the NSTDA of research topics from agricultural science to centers, operating outside the normal framework of biomedical science and environmental science. In civil service and state enterprises. This enabled the addition to research units, development units have Center to operate more effectively to support and been established for activities with high commercial transfer technology for the development of industry, potential. These are full scale business and agriculture, natural resources, environment and production operations designed to demonstrate the consequently the social and economic well-being commercial viability of technologies to prospective of Thai people. Other centers under the NSTDA investors. family include National Metal and Materials Technology Center (MTEC), National Electronics and Apart from research and commercialization, BIOTEC Computer Technology Center (NECTEC), National activities also include policy research, an outreach Nanotechnology Center (NANOTEC) and Technology program, human resource development and Management Center (TMC). international relations. 8 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Organization
BIOTEC Research Units at Thailand Science Park • Bioresources Technology Research Unit • Animal Biotechnology Research Unit • Food Biotechnology Research Unit • Medical Molecular Biology Research Unit • Genome Technology Research Unit • Biosensing Technology Research Unit
Collaborative Research Laboratories at universities and government organization • Biochemical Engineering and Pilot Plant Research and Development Laboratory - at King Mongkut’s University of Technology Thonburi (KMUTT) • Waste Utilization and Management Laboratory - at King Mongkut’s University of Technology Thonburi (KMUTT) • Cassava and Starch Technology Research Laboratory - at Kasetsart University • Rice Gene Discovery Laboratory - at Kasetsart University • Medical Biotechnology Research Laboratory - at Faculty of Medicine Siriraj Hospital and Chiang Mai University • Biomedical Technology Research Laboratory - at Chiang Mai University • Marine Biotechnology Laboratory - at Chulalongkorn University • Molecular Biology and Genomics of Shrimp Laboratory - at Chulalongkorn University • Shrimp Molecular Biology and Biotechnology Laboratory - at Mahidol University • Peat Swamp and Hala-Bala Rain Forest Research Laboratory - jointly established with the National Park, Wildlife and Plant Conservation Department and located in Narathiwat Province
Translational Administration Research Facilities and Multi-disciplinary • Policy Study and Biosafety Unit Laboratory • Rural Development Technology Service Unit • Biotechnology Business Development Division • Shrimp Genetic Improvement Center • Human Resources and Platform Technology • Nuclear Polyhedrosis Virus Pilot Development Division Plant for Insect Pest Control • Strategic Planning and Organization • Thailand Bioresource Research Center Development Division • Food and Feed Innovation Center • Evaluation and Monitoring Division • Integrative Biorefinery Laboratory • Research Unit Coordination Division • National Biopharmaceutical Facility • Management Information System Division (in collaboration with King Mongkut’s • International Cooperation and Public University of Technology Thonburi) Relations Division • Building Management and Equipment Service Division • General Management Division Facts and Figures 9
Expenditure, 775.7 m THB Human Resources, 579 Technology Transfer, 3% Research Management, 3% Human Resource Development, 4% Platform Technologies, 11% Infrastructure, 8% Cross-cutting Technologies, 0% 26 Internal Management,15% Bioresources and Community, 22% (5%) 3% Energy and Environment, 1% 4% 3% Health and Medicine, 9% 8% Agriculture and Food, 24% 158 (27%) 180 (31%) 11%
15%
22% 215 (37%) 24% 1% 9% PhD., 31% B.Sc., 27% M.Sc., 37% Below B.S., 5%
Socio-economic impact Income from sources outside NSTDA, 107.74 m THB of 5,270 m THB 2% generated by 49 1% completed project 1% F 15% Investment generation Research Fund 62.73 m THB 127 Contract/Collaborative m THB 107.74 58% Research 24.84 m THB m THB Technical Services 16.10 m THB Revenue generation 23% Workshop/Conference 1.32 m THB 4,540 Licensing Activity 1.34 m THB m THB Others 1.41 m THB
Cost reduction 509 m THB Import replacement Major Outputs 91 m THB Intellectual Properties
granted patent overseas Publications 1 233 papers, including 16 granted patent in Thailand papers in non-citation index 1 journals 30 granted petty patents in Thailand 27 patent applications in Thailand Honors and Awards 12 awards and honors 23 petty patent applications in Thailand 5,270 m THB 2 trade secret applications in Thailand 10 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Research and Development 11
RESEARCH AND FDEVELOPMENT
BIOTEC’s R&D and technical program covers a wide range of topics. Plant biotechnology focuses on three economically important plants: rice, cassava, and oil palm. Animal biotechnology focuses on shrimp and dairy cows, whereas food biotechnology aims to improve and upgrade the processing and quality of fermented food, including topics such as food safety and risk assessment, food chemistry, and starter culture technology. Medical biotechnology focuses on tropical and emerging diseases such as malaria, tuberculosis, dengue fever and influenza. On environmental issues, BIOTEC gives emphasis to the study of microbial diversity and the preservation, use and conservation of bioresources. Biogas and other renewable energies are the focus in the energy research theme. 12 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Plant and grain types of pseudo-backcross inbred lines PinK + 4 (A-C) Morphology of paddy and milled rice grains compared with pseudo-recurrent parent PinK3 (D). of Myanmar Paw San Hmwe accessions.
Ref: Ruengphayak, S., Chaichumpoo, E., Phromphan, S., Kamolsukyunyong, Highlights from Rice Biotechnology W., Sukhaket, W., Phuvanartnarubal, E., Korinsak, S., Korinsak, S., Vanavichit, A. (2015). Pseudo-backcrossing design for rapidly Using pseudo-backcrossing scheme to expedite pyramiding multiple traits into a preferential rice variety. Rice, 8:7 rice gene-pyramiding Genetic characterization of the world’s best rice, Rice production in irrigated areas of Thailand has been Myanmar Paw San Hmwe frequently and strongly affected by abiotic stresses resulting from unfavorable climatic changes, such as Paw San Hmwe (PSM) rice from Myanmar is among flooding and drought, as well as by biotic stresses the best rated rice varieties. It was named the World caused by bacterial leaf blight (BB), leaf/neck blast Best Rice 2011. It is cultivated in many areas of (BL) and brown planthopper (BPH). Therefore, new Myanmar. Strong aroma, good taste and its elongation successful breeding lines must possess multiple types during cooking are its key characteristics. However, of resistance to both biotic and abiotic stresses, as the molecular marker-based analysis of prominent well as demonstrating specific grain qualities and high traits and genetic/phenotypic characteristics in the yield. However, pyramiding multiple genes into a Paw San rice has not been well studied. desirable genetic background can take years to accomplish. The genetic characterization of PSM accessions obtained from the Myanmar germplasm bank was To shorten the time, researchers redesigned the gene- investigated. Thirty-one PSM accessions were pyramiding platform by integrating marker assisted genotypically characterized and their physical grain selection into pseudo-backcross breeding. With this and cooking quality traits were studied. Specific gene platform, they were successful in pyramiding five markers associated with aroma, apparent amylose functional genes (xa5, Xa21, Sub1A-C, SSIIa, TPS) and content (AAC) and alkali spreading value were used three QTLs (qBph3, qBL1, qBL11) into the ‘PinK3’ to determine the alleles carried by different PSM genome background within only seven breeding cycles accessions. The characterization and grouping data in four years. ‘PinK3’ is an aromatic, high-yielding, of PSM accessions posted benefits to Myanmar seed non-photoperiod-sensitive, high-amylose rice variety, banks, and these results can facilitate the utilization but it is susceptible to BPH, BB, BL and submergence of PSM rice as a genetic resource in rice breeding stresses. The new, improved lines have a high-yield programs. phenotype that confers submergence tolerance and resistance to BPH, BB and BL. This is the first report This study was jointly conducted by researchers from describing the application of pseudo-backcrossing to Rice Gene Discovery Laboratory, Kasetsart University, significantly shorten the time required for gene/QTL Myanmar Department of Agricultural Research and pyramiding in an annual crop (rice). Myanmar Agriculture Service.
This work was a collaborative effort between Rice ,Ref: Oo K.S., Kongjaimun, A., Khanthong, S., Yi, M., Myint, T.T., Korinsak, S., Siangliw, J.L., Myint, K.M., Vanavichit, A., Malumpong, C. Gene Discovery Laboratory and Kasetsart University. and Toojinda T. (2015). Characterization of Myanmar Paw San Hmwe Accessions Using Functional Genetic Markers. Rice Science, 22(2), 53-64. Research and Development 13
Insight into brown planthopper resistance To find an alternative to chemical bacteriocides, mechanisms in Thai Jasmine rice scientists explored the use of bacteriophages as biological control agents for R. solanacearum. In Thailand, brown planthopper (BPH; Nilaparvata Fourteen phages infecting R. solanacearum were lugens) infestation is one of the major annual disease isolated from soil samples collected in tomato fields outbreaks in rice crops, causing huge cultural and in Chiang Mai, Thailand. The phages showed different economic burdens to Thai Jasmine rice producers. host ranges when tested against 59 R. solanacearum Khao Dawk Mali 105 (KDML 105; KD), a well-known strains isolated from Thailand and Japan. These Thai Jasmine rice, is highly susceptible to BPH. phages were characterized as nine podoviruses and Planthoppers damage rice directly through feeding five myoviruses based on their morphology. The and also by transmitting two viruses, rice ragged stunt podoviruses isolated in this study showed strong lytic virus and rice grassy stunt virus. Up to 60% yield loss activity and wide host ranges. Therefore, these phages is common in susceptible rice cultivars attacked by BPH. have potential use in the decontamination of pathogen-infected field soils. Phage J2 demonstrated To better understand the biological mechanism of an ability to prevent bacterial wilt of tomato in pot infestation, researchers investigated the metabolomic experiments. responses to BPH infestation in Thai rice varieties.1 H NMR spectroscopy, combined with chemometrics, This study was a collaborative effort between Genome was used to analyze the polar metabolome from leaf Technology Research Unit, Kasetsart University and extracts of Thai Jasmine rice and its BPH resistant Hiroshima University (Japan). isogenic lines with and without BPH infestation at
various time points. The study identified, for the first Ref: Bhunchoth, A., Phironrit, N., Leksomboon, C., Chatchawankanphanich, time, several potential metabolic pathways for O., Kotera, S., Narulita, E., Kawasaki, T., Fujie, M. and Yamada, T. acclimatization and defense mechanisms against BPH (2015) Isolation ofRalstonia solanacearum-infecting bacteriophages from tomato fields in Chiang Mai, Thailand, and their experimental use infestation. These findings provide a valuable, first as biocontrol agents. Journal of Applied Microbiology, 118(4), 1023-33. insight into BPH resistance mechanisms in Thai Jasmine rice. Large-scale SNP discovery in cassava via F transcriptome sequencing This study was conducted by researchers from BIOTEC, Cassava is one of the most important crop species, Kasetsart University, Queen’s University Belfast (UK) having multiple uses including starch and being the and Beaumont Health System (USA). main source of dietary energy in several less developed Ref: Uawisetwathana, U., Graham, S.F., Kamolsukyunyong, W., Sukhaket, countries. Marker-assisted selection has become an W., Klanchui, A., Toojinda, T., Vanavichit, A., Karoonuthaisiri, N. and essential tool in plant breeding. Single nucleotide Elliott, C.T. (2015). Quantitative 1H NMR metabolome profiling of Thai Jasmine rice (Oryza sativa) reveals primary metabolic polymorphism (SNP) discovery via transcriptome response during brown planthopper infestation. Metabolomics, sequencing is an attractive strategy for genome 11, 1640-1655. complexity reduction in organisms with large genomes.
Highlights from Plant and Animal Biotechnology
The use of bacteriophage to control bacterial wilt Bacterial wilt caused by Ralstonia solanacearum is one of the most devastating diseases of many economically important crops in Thailand such as ginger, pepper, tomato, potato and Curcuma alismatifolia Gagnep. At present, protection from losses by bacterial wilt is achieved mainly by early detection and subsequent eradication by destroying
the host, usually by using chemical bacteriocides. Cassava is a major staple food in the developing world, as well as a source for producing ethanol. 14 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Researchers sequenced the transcriptome of 16 placed on the genetic map. This SNP-based linkage cassava accessions using the Illumina HiSeq platform map was subsequently employed in a QTL analysis to and identified 675,559 EST-derived SNP markers. A detect markers associated with fruit bunch weight subset of those markers was subsequently genotyped and trunk height. These markers will be useful for by capture-based targeted enrichment sequencing in selecting individual palms with desirable characteristics
100 F1 progeny segregating for starch viscosity in breeding programs. Furthermore, the high-density phenotypes. A total of 2,110 non-redundant SNP map will contribute to a fundamental knowledge of markers were used to construct a genetic map. This genome structure and will be valuable for mapping map encompasses 1,785 cM and consists of 19 linkage other economically important genes for marker- groups. A major quantitative trait locus (QTL) assisted selections. controlling starch viscosity was identified and shown to coincide with the QTL previously reported for this This study was performed by Genome Technology trait. This work represents the first effort to perform Research Unit. capture-based targeted enrichment sequencing in cassava and illustrates the attractiveness of this Ref: Pootakham, W., Jomchai, N., Ruang-Areerate, P., Shearman, J.R., Sonthirod, C., Sangsrakru, D., Tragoonrung, S approach for genotyping SNPs in predetermined and Tangphatsornruang, S. (2015). Genome-wide SNP discovery and genomic regions. identification of QTL associated with agronomic traits in oil palm using genotyping-by-sequencing (GBS). Genomics, 105(5-6), 288-95. This study was a collaborative effort by researchers Finding genes responsible for oil palm yield from Mahidol University and Genome Technology Research Unit. Oil palm (Elaeis guineensis), the tropical perennial and cross-pollinated crop, is the most productive oil- Ref: Pootakham, W., Shearman, J.R., Ruang-Areerate, P., Sonthirod, producing crop. Improvement of oil palm yield could C., Sangsrakru, D., Jomchai, N., Yoocha, T., Triwitayakorn, K., Tragoonrung, S. and Tangphatsornruang, S. (2014). Large-scale significantly contribute to the overall oil production SNP discovery through RNA sequencing and SNP genotyping by in the world. The number of fruit bunches is one yield targeted enrichment sequencing in cassava (Manihot esculenta Crantz). PLOS ONE, 9(12):e116028. doi: 10.1371/journal. component that can be improved by increasing sex pone.0116028. ratio, the ratio of female inflorescences to total inflorescences. Genome-wide SNP discovery in oil palm using genotyping-by-sequencing approach Researchers attempted to identify gene(s) by making a physical map of a specific locus responsible for sex With rapid advancement in sequencing throughput, ratio. A putative aldo-keto reductase gene together with an overall decrease in sequencing cost, (named EgAKR1) was revealed to be a promising next generation sequencing technologies have been candidate for sex ratio determination, via controlling applied to SNP identification in various plant species. female inflorescence number. This was predicted from However, it remains costly to employ whole-genome the two newly identified polymorphic marker loci sequencing to evaluate multiple individuals in a (mEgSSRsr8-21LB and mEgAKR1-9) designed from mapping population, especially for organisms with EgAKR1. The functions of AKR gene families in other large genomes such as oil palm. Reduced representation plant species and the promoter analysis suggested methods are extremely useful, not only because of that EgAKR1 may contribute to the sex ratio through their cost-reducing aspects, but also because many abiotic stress responsiveness. research questions can be answered with a small set of markers and do not require every base of the This investigation was made by researchers from genome to be sequenced. Genotyping-by-sequencing Genome Technology Research Unit and Thammasat (GBS) is an efficient strategy that can simultaneously University. detect and score tens of thousands of molecular markers. Ref: Somyong, S., Poopear, S., Jomchai, N., Uthaipaisanwong, P., Ruang- areerate, P., Sangsrakru, D., Sonthirod, C., Ukoskit, K., Tragoonrung, Researchers employed GBS approach to perform a S. and Tangphatsornruang, S. (2015). The AKR gene family and modifying sex ratios in palms through abiotic stress large-scale SNP discovery and genotyping of a responsiveness. Functional & Integrative Genomics, 15(3), 349-362. mapping population in oil palm. Over 21,000 SNP markers were identified and 1085 markers were Research and Development 15
generating a common set of genome-wide SNP data suitable for constructing integrated linkage maps from multiple populations in a highly heterozygous agricultural species.
This was a collaborative work between Genome Technology Research Unit and Rubber Research Institute of Thailand.
Ref: Pootakham, W., Ruang-Areerate, P., Jomchai, N., Sonthirod, C., Sangsrakru, D., Yoocha, T., Theerawattanasuk, K., Nirapathpongporn, K., Romruensukharom, P., Tragoonrung, S. and Tangphatsornruang, S. (2015). Construction of a high-density integrated genetic linkage map of rubber tree (Hevea brasiliensis) using genotyping-by- sequencing (GBS). Frontiers in Plant Science, 6, 367.
GBS-based genetic linkage maps of rubber tree derived from F1 progeny from (A) BPM24 × RRIM600 and (B) BPM24 × RRIC110 crosses.
F Porcine epidemic diarrhoea virus has a substantial economic burden High-density integrated genetic linkage map of given that it is highly infectious, resulting in significant morbidity and mortality in piglets. rubber tree Construction of linkage maps is crucial for genetic studies and marker-assisted breeding programs. Development of porcine epidemic diarrhoea Recent advances in next generation sequencing vaccine technologies allow for the generation of high-density Porcine epidemic diarrhoea virus (PEDV) causes linkage maps, especially in non-model species lacking acute diarrhoea and dehydration in swine of all ages, extensive genomic resources. with significant mortality in neonatal pigs. The recent rise of PEDV outbreaks in Asia and North America Researchers employed the genotyping-by-sequencing warrants an urgent search for effective vaccines. (GBS) technique to perform a genome-wide SNP However, PEDV vaccine research has been hampered discovery and genotyping of two rubber tree mapping by difficulties in isolating and propagating the virus populations. Single-population linkage maps were in mammalian cells, thereby complicating the generated and common SNP markers were used as recovery of infectious PEDV using a full-length bridges to merge them into a high-density integrated infectious clone. genetic map, possibly the most saturated genetic map on rubber tree to date. SNP markers reported in this In this study, researchers engineered VeroE6 cells to study will expand the existing repertoire of available stably express porcine aminopeptidase N (pAPN) and molecular markers in rubber tree, and the integrated used them as a platform to obtain a high-growth
genetic map presented will be useful for future variant of PEDV, termed PEDVAVCT12. Subsequently, breeding programs, association studies with desirable the full-length cDNA clone was constructed by agronomic traits, genetic diversity analyses and assembling contiguous cDNA fragments
phylogenetic studies. This study also demonstrated encompassing the complete genome of PEDVAVCT12 that GBS is a robust and cost-effective approach for 16 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
in a bacterial artificial chromosome. Infectious PEDV Researchers investigated PmFAMeT gene which is could be recovered, and the rescued virus displayed believed to regulate growth and reproduction of P. phenotypic properties identical to the parental virus. monodon. Two full-length cDNAs of PmFAMeT were
Interestingly, PEDVAVCT12 was found to contain a identified, in order to examine molecular involvement C-terminal deletion of the spike gene, resulting in of PmFAMeT gene products on ovarian (and oocyte) disruption of the ORF-3 start codon. When a functional development. PmFAMeT was found to be differentially ORF-3 gene was restored, the recombinant virus could expressed during ovarian development of P. monodon. not be rescued, suggesting that ORF-3 could suppress Eyestalk ablation and exogenous serotonin injection PEDV replication in vitro. In addition, a high-growth can promote PmFAMeT expression. and genetically stable recombinant PEDV expressing a foreign protein could be rescued by replacing the In a separate study, researchers evaluated biological ORF-3 gene with the mCherry gene. Together, the roles of X-box binding protein 1 in reproduction and results of this study provide a means to generate growth of P. monodon by characterizing genetically defined PEDV as a promising vaccine the PmXbp1 cDNA sequence. Expression patterns of candidate. PmXbp1 during ovarian development in wild P. monodon broodstock were examined. The study This study was conducted by Animal Biotechnology showed that eyestalk ablation had an effect on the Research Unit. expression level of PmXbp1 during late ovarian development, whereas serotonin injection promoted Ref: Jengarn, J., Wongthida, P., Wanasen, N., Frantz, P.N., Wanitchang, the expression level of ovarian PmXbp1. A. and Jongkaewwattana, A. (2015) Genetic manipulation of porcine epidemic diarrhoea virus recovered from a full-length infectious cDNA clone. Journal of General Virology, 96(8), 2206-18. Both studies were conducted by Animal Biotechnology Research Unit in collaboration with Chulalongkorn University. Highlights from Shrimp Biotechnology Ref: 1. Buaklin, A., Jantee, N., Sittikankaew, K., Chumtong, P., Janpoom, S., Study of shrimp growth and reproduction Menasveta, P., Klinbunga, S. and Khamnamtong, B. (2015). Expression and polymorphism of farnesoic acid O-methyltransferase (FAMeT) Closing life cycle culture is crucial to the domestication and association between its SNPs and reproduction-related parameters of the giant tiger shrimp Penaeus monodon. Aquaculture, and genetic improvement of the giant tiger shrimp 441, 106-117. (Penaeus monodon). However, poor reproductive 2. Prasertlux, S., Yocawibun, P., Janpoom, S., Klinbunga, S., Menasveta, P. maturation of captive P. monodon females and low and Khamnamtong, B. (2015). Differential expression of X-box binding quality of spermatozoa of captive males have limited protein 1 during ovarian development and association between its SNP and growth-related parameters of the giant tiger shrimp Penaeus the potential of genetic improvement, which in turn, monodon. Aquaculture, 448, 531-538. resulted in remarkably slow domestication and selective breeding programs of P. monodon in Transmission of yellow head virus Thailand. Yellow head virus (YHV) is an economically important disease in farmed shrimp in South East Asia. Despite the continual search since its discovery in the early 1990s, the reservoir for YHV-1, the most virulent strain of YHV found in Thailand, has still not been identified. Outbreaks of the virus in cultivated, exotic white leg shrimp P. vannamei that originate from SPF stocks known to be free of the virus, suggested that the outbreaks occur via horizontal transmission from an environmental source. Since the Australian red claw crayfish, Cherax quadricarinatus and the marine penaeid shrimp are often cultivated in adjacent areas Examples of red claw crayfish and shrimp gill tissues immuno-stained in Thailand, it is of importance to know whether red with monoclonal antibodies against YHV nucleocapsid protein and claw crayfish is susceptible to YHV and whether YHV counter stained with hematoxylin and eosin. could be transmitted between these species. Research and Development 17
Researchers performed the tests whether red claw Ref: Rungrassamee, W., Kingcha, Y., Srimarut, Y., Maibunkaew, S., Karoonuthaisiri, N. and Visessanguan, W. (2014). crayfish was susceptible to endemic YHV and also Mannooligosaccharides from copra meal improves survival of the capable of transmitting it to black tiger shrimp. The Pacific white shrimp (Litopenaeus vannamei) after exposure to Vibrio results revealed that red claw crayfish is susceptible harveyi. Aquaculture, 434, 403–410. but highly tolerant to the virus. Infected red claw crayfish could transmit the virus without showing any signs of yellow head disease. These facts make them Highlights from Food Science and a perfect carrier for the virus. Biotechnology
This study was conducted by researchers from BIOTEC Discovery of a halotolerant yeast, a potential Shrimp Molecular Biology and Biotechnology starter culture for soy sauce fermentation Laboratory, Mahidol University, Institute of Veterinary Soy sauce is made by the fermentation of soybeans Research and Development of Central Vietnam and combined with wheat flour, rice flour and brine. The Institute of Biotechnology (Vietnam). production involves two steps: koji fermentation using Aspergillus oryzae and moromi fermentation by Ref: Soowannayan, C., Nguyen, G.T., Pham, L.N., Phanthura, M. and Nakthong, N. (2015). Australian red claw crayfish (Cherax adding brine solution into the koji. In moromi quadricarinatus) is susceptible to yellow head virus (YHV) infection fermentation, glutaminase produced by naturally- and can transmit it to the black tiger shrimp (Penaeus monodon). present yeast converts l-glutamine originated from Aquaculture, 445, 63-69. soy protein to l-glutamic acid, a compound responsible for “umami” taste. Given the high salinity condition Immunostimulant effect of copra meal MOS on of moromi fermentation, a halotolerant yeast would white shrimp be beneficial to the soy sauce fermentation. Mannooligosaccharides (MOS) are linear chains of mannose sugars. They are known to possess prebiotic Researchers investigated yeast isolates obtained from affect and therefore can be used as a feed additive to Thai soy sauce fermentation. The most interesting prevent pathogen colonization and modulate the yeast was identified as Meyerozyma (Pichia) Fimmune system of host animals. Most MOS used in guilliermondii EM2Y61. This strain is a salt-tolerant animal feed additives are derived from the cell wall yeast that could tolerate up to 20% (w/v) NaCl and of yeast. However, copra meal, a waste product from produce extracellular and cell-bound glutaminases. coconut industries, is another rich source for MOS. The extracellular glutaminase activity was found to be much higher than that of cell-bound glutaminase. BIOTEC researchers investigated the effects of MOS This is the first report of glutaminase producing M. from copra meal as a dietary feed additive on growth guilliermondii isolated from the moromi of Thai soy performance, disease resistance, and immune sauce fermentation. M. guilliermondii EM2Y61 has enhancement of the Pacific white shrimp Litopenaeus high potential to be developed into starter yeast vannamei. The findings showed that MOS from copra culture to increase l-glutamic acid during soy sauce meal was able to significantly increase protection in fermentation. the Pacific white shrimp upon pathogen exposure, while it had no significant effect on shrimp growth This was a joint study between Mahidol University rate and feed conversion ratio. Moreover, MOS and Food Biotechnology Research Unit. supplementation induced crucial shrimp immune- related genes associated with antimicrobial peptides and the peritrophic membrane. The findings suggested that MOS can potentially be applied as an immunostimulant, particularly in aquaculture, and copra meal, which is considered a waste product of the coconut industry, can be an alternative source for MOS.
This study was conducted by researchers from Food Biotechnology Research Unit and Biosensing Technology Research Unit. Cassava pulp, waste from starch factory, can be a potentially good source of prebiotics, because of its high hemicelluloses content that promotes the growth of gut flora. 18 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Ref: Aryuman, P., Lertsiri, S., Visessanguan, W., Niamsiri, N., Bhumiratana, Researchers isolated a phage from an infected culture A. and Assavanig, A. (2015). Glutaminase-producing Meyerozyma (Pichia) guilliermondii isolated from Thai soy sauce fermentation. of B. amyloliquefaciens FB11, a strain used locally in International Journal of Food Microbiology. 192, 7-12. Thailand. The phage was classified morphologically into the Caudovirales order, Myoviridae family. In Novel bacteriocins: Natural antimicrobial agents order to eliminate phage infection, an early detection for food and feed industry of the phage and an inactivation measure must be applied in combination. Researchers therefore Contamination by food-borne pathogens is one of the developed a PCR-based method that can detect the major problems in food industry. Among the techniques phage with a 104 PFU/mL limit of detection, in less used to control the microbial contamination in food than 3 h including sample treatment, PCR analysis and is the use of natural antimicrobial agents. Bacteriocins gel electrophoresis. The study also demonstrated that or antimicrobial peptides produced by lactic acid the phage could be inactivated by either thermal bacteria (LAB) hold promise as natural antimicrobial treatment at 70°C for 5 min or treatment with agents. peracetic acid-based disinfectant (0.3 % v/v) for 5 min. Researchers discovered two small novel bacteriocins, This work was conducted by Food Biotechnology 7293A and 7293B, from Weissella hellenica BCC 7293, Research Unit. isolated from Thai fermented pork sausage. Both
bacteriocins had broad antimicrobial spectra and Ref: Pitaksutheepong, C., Abhisingha, M., Dumnin, J. and Visessanguan, exceptionally inhibited several important Gram- W. (2015). Isolation, detection and inactivation of a Myoviridae negative food-borne pathogens including bacteriophage infecting Bacillus amyloliquefaciens FB11. Annals of Microbiology, DOI 10.1007/s13213-014-1022-9. Pseudomonas aeruginosa, Aeromonas hydrophila, Salmonella Typhimurium and Escherichia coli. The Prebiotics from rice bran and cassava pulp sensitivity to proteolytic enzymes and stability in organic solvents, pH and temperature of both Agricultural wastes such as rice bran and cassava pulp bacteriocins suggested the potential use of bacteriocin are inherently rich in plant cell wall materials like 7293A and B in food and feed applications, replacing hemicelluloses, which are complex heteropolymers antibiotics. consisting of several kinds of monosaccharides connected by various glycosidic linkages. Due to this This study was a collaborative work between Food variety of glycosidic bonds between the monomers, Biotechnology Research Unit, Prince of Songkla hemicelluloses are usually not digestible by human University and Kyushu University (Japan). intestinal digestive enzymes, but could be utilized by members of the intestinal microbiota. The fact that Ref: Woraprayote, W., Pumpuang, L., Tosukhowong, A., Roytrakul, S., hemicelluloses can be utilized by some beneficial Perez, R.H., Zendo, T., Sonomoto, K., Benjakul, S. and Visessanguan, W. (2015). Two putatively novel bacteriocins active against Gram- microbes present in the intestine makes hemicellulose- negative food borne pathogens produced by Weissella hellenica BCC containing substances potential prebiotic sources. 7293. Food Control, 55, 176-184. Researchers investigated the prebiotic potential of Eliminating phage infection in animal feed rice bran and cassava pulp. Hydrothermal treatments production were used to extract hemicellulosic oligosaccharide Fermented soybean meal is commonly used as a mixtures from rice bran and cassava pulp. Microbial protein source in animal feed, with Bacillus utilization of the obtained oligosaccharide mixtures amyloliquefaciens primarily used as a starter culture showed that they were able to promote the growth for fermentation. The fermentation industry considers of two from three Lactobacillus, as well as three from phage attacks a major threat as they can cause serious five Bifidobacterium species tested. From the three yield losses. Thus, characterization and inactivation tested Bacteriodes strains, one utilized the cassava of these phages would be essential in managing phage pulp oligosaccharide mixture better than inulin, while decontamination in fermented feed production two grew better on rice bran oligosaccharide mixture factories. than on inulin. Furthermore, the two oligosaccharide mixtures were found to be stable at 85°C for 30 min. Similarly, these mixtures were able to withstand their exposure to simulated human gastric juice (pH 1–5) Research and Development 19
and to pancreatin treatments for up to two hours. Molecular characterization of Plasmodium These findings suggested that rice bran and cassava falciparum Bruno/CELF RNA binding proteins. pulp can be a potentially good source of prebiotics. The human malaria parasite Plasmodium falciparum employs intricate post-transcriptional regulatory This research was a joint study between Mahidol mechanisms in different stages of its life cycle. Despite University and Food Biotechnology Research Unit. the importance of post-transcriptional regulation, key
Ref: Kurdi, P. and Hansawasdi, C. (2015). Assessment of the prebiotic elements of these processes, namely RNA binding potential of oligosaccharide mixtures from rice bran and cassava proteins (RBPs), are poorly characterized. pulp. Lwt-Food Science and Technology, 63(2), 1288–1293. Researchers characterized the RNA binding properties of P. falciparum proteins, including two putative Highlights from Malaria Research members of the Bruno/CELF family of RBPs (PfCELF1 and PfCELF2), dihydrofolate reductase-thymidylate Bacterial surrogate system: A tool to explore synthase (PfDHFR-TS), and adenosine deaminase antimalarial drug interaction (PfAda). RNA binding activity was tested using UV- With continuing global threat of malaria, there is an crosslinking and electrophoretic mobility shift assays. urgent need to search not only for new drugs, but also PfCELF1 and PfDHFR-TS demonstrated RNA binding for effective drug combinations. Inhibitors of activity, whereas PfAda and PfCELF2 were RBP- dihydrofolate reductase (DHFR) such as pyrimethamine negative. Intracellular protein localization of RBPs was and of dihydropteroate synthase (DHPS) such as sulfa studied using GFP-tagged transgenic parasite drugs are known to have synergistic interactions. lines. PfCELF1 protein may shuttle between nucleus However, studies of the synergism are complicated and cytoplasm, as shown by a predominantly by the fact that the malaria parasite can also salvage nuclear PfCELF1 cell population and another exogenous folates, and the salvage may also be predominantly cytoplasmic. In contrast, PfDHFR-TS affected by the drugs. It is desirable to have a protein is predominantly cytoplasmic. PfCELF1 may convenient system to study interaction of DHFR and thus have several roles, including pre-mRNA FDHPS inhibitors without such complications. processing. The mRNA targets of these P. falciparum proteins were investigated by ribonomics Researchers used Escherichia coli transformed with using DNA microarrays. A sequence motif similar to malarial DHFR and DHPS, while its own corresponding that recognized by CELF proteins in other species is genes were inactivated by optimal concentration of common in the introns of target mRNAs identified for trimethoprim and genetic knockout, respectively, to PfCELF1, suggesting that nuclear-localized PfCELF1 study the interaction of the inhibitors. Marked may regulate pre-mRNA splicing in P. falciparum, as synergistic effects are observed for all combinations has been found for CELF proteins in other species. In of pyrimethamine and sulfa inhibitors in the presence contrast, none or very few mRNA targets were found of trimethoprim. The results show synergism between for the other proteins, suggesting that they do not inhibitors of the two enzymes even in the absence of have biologically relevant roles as RBPs in the asexual folate transport and uptake. This bacterial surrogate stages of P. falciparum. system can, therefore, be used as a tool for assessing the interactions of drug combinations between the This work was conducted by Medical Molecular DHFR and DHPS inhibitors. Biology Research Unit in collaboration with the University of Edinburgh. This study was conducted by researchers from Mahidol University and Medical Molecular Biology Ref: Wongsombat, C., Aroonsri A., Kamchonwongpaisan, S., Morgan, H.P., Walkinshaw, M.D., Yuthavong, Y. and Shaw P.J. (2014). Molecular Research Unit. characterization ofPlasmodium falciparum Bruno/CELF RNA binding proteins. Molecular and Biochemical Parasitology, 198(1), 1-10. Ref: Talawanich, Y., Kamchonwongpaisan, S., Sirawaraporn, W. and Yuthavong, Y. (2015). Use of bacterial surrogates as a tool to explore antimalarial drug interaction: Synergism between inhibitors of malarial dihydrofolate reductase and dihydropteroate synthase. Acta Tropica, 149, 64-69. Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
r stal structure o Plasmodium vivax se rine netic ec an s o Plasmodium vivax se rine ro et ltrans erase ro et ltrans erase Plasmodium ara ite the ca ati e agent of alaria Plasmodium T ha een ho n to e e ential rely hea ily on de novo folate io ynthe i and the for ara ite gro th and de elo ent a ing it a en y e in thi ath ay ha e therefore een e lored ri e target for anti alarial dr g che othera y e ten i ely for anti alarial de elo ent erine de elo ent n in de th nder tanding of hydro y ethyltran fera e ( T) fro Plasmodium Plasmodium T inetic and echani i needed an en y e in ol ed in folate recycling and dT o that the di erence in reaction detail a ong ynthe i ha een ho n to cataly e the con er ion T can e nder tood and ed for the of and erine to glycine (Gly) in a T de endent de elo ent of eci c inhi itor reaction the echani of hich i not yet f lly nder tood e earcher e loyed ario ethodologie incl ding ligand inding ea re ent a ell a In thi t dy re earcher deter ined the cry tal tran ient and teady tate inetic to in e tigate the tr ct re of Plasmodium vivax T (Pv T) in a T reaction The re lt gge ted that the inary co le ith erine and in a ternary co le inetic echani of T occ r ia a rando ith erine ( er) and ( R) for yl tetra hydro order odel and that glycine for ation i the rate folate ( T ) hich gge t the echani li iting te of T Thi infor ation ill er e a nderlying the control of en y e acti ity The re lt a a i for f t re in e tigation for other T The a ert the i ortance of feat re ch a co arati e t die of T fro ara ite and ho t tereo electi ity and redo tat for control of the ill a e the ay for the de ign of ecie eci c acti ity and eci city of Pv T The tr ct re inhi itor for anti alarial dr g de elo ent f nction analy i of Pv T ha led to the de ign of anti alarial targeting T Thi or a a colla orati e e ort et een re earcher fro ahidol ni er ity B ra ha Thi or a a colla orati e e ort et een ni er ity Ch lalong orn ni er ity the ni er ity re earcher fro ahidol ni er ity B ra ha of ichigan ( ) edical olec lar Biology ni er ity National ynchrotron adiation e earch e earch nit and Bio en ing Technology e earch Center (Tai an) edical olec lar Biology e earch nit nit and Bio en ing Technology e earch nit e aen en orn atchara ong ra atong charita l e Chitn ar at iangr ngro I arat Noytano alfey B tha ong Chitn eart a l anich Oonanant anichthanan l Ch an hayan aen en and Chaiyen ( ) inetic echani and the rate li iting te Plasmodium vivax Journal of Chen C Chaiyen tha ong and eart a l anich ( ) of erine hydro y ethyltran fera e Biological Chemistry tr ct re of Plasmodium vivax erine hydro y ethyltran fera e ( ) i lication for ligand inding eci city and f nctional control Acta Crystallographica Section: D Biological Crystallography ( t ) l ts ro en ue esearc
ole o ost prote n n en ue rus repl cation o t and iral rotein are in ol ed in deng e ir ( EN ) re lication eterogeneo ri on cleo rotein (hn N ) C C are a ndant ho t cell lar rotein that e hi it N inding acti ity and lay i ortant role in the re lication of o iti e trand N ir e ch a olio ir and he atiti C ir hn N C C ha e re io ly een ho n to interact ith i entin and iral N in EN infected cell ho e er their f nctional role in EN re lication i not clearly Cry tal tr ct re of Plasmodium vivax erine hydro y ethyltran fera e (Pv T) nder tood Research and Development 21
Researchers investigated the role of hnRNP C1/C2 in Ref: Yasamut, U., Tongmuang, N., Yenchitsomanus, P., Junking, M., Noisakran, S., Puttikhunt, C., Chu, J.J. and Limjindaporn, T. (2015). DENV replication by using an in vitro model of DENV Adaptor Protein 1A Facilitates Dengue Virus Replication.PLOS ONE, infection in a hepatocyte cell line (Huh7) and siRNA- 10(6): e0130065. doi:10.1371/journal.pone.0130065. mediated knockdown of hnRNP C1/C2. The results suggest that hnRNP C1/C2 is involved in DENV Potential biomarker for prediction of severe replication at the stage of viral RNA synthesis. These dengue findings pave the way for further study on the Shedding of microparticles (MPs) is a consequence of molecular mechanisms of the viral and host protein apoptotic cell death and cellular activation. Low levels interactions required for viral replication in DENV- of circulating MPs in blood help maintain homeostasis, infected cells. whereas increased MP generation is linked to many pathological conditions. This study was conducted by Chiang Mai University; Faculty of Medicine Siriraj Hospital, Mahidol University, In this study, researchers investigated the role of MPs Medical Biotechnology Research Laboratory and in dengue virus (DENV) infection. Infection of various Biomedical Technology Research Laboratory. susceptible cells by DENV led to apoptotic death and
Ref: Dechtawewat, T., Songprakhon, P., Limjindaporn, T., Puttikhunt, MP release. These MPs harbored a viral envelope C., Kasinrerk, W., Saitornuang, S., Yenchitsomanus, P. and Noisakran, protein and a nonstructural protein 1 (NS1) on their S. (2015). Role of human heterogeneous nuclear ribonucleoprotein surfaces. Ex vivo analysis of clinical specimens from C1/C2 in dengue virus replication. Virology Journal, 12, 14. patients with infections of different degrees of severity at multiple time points revealed that MPs generated Adaptor protein 1A facilitates dengue virus from erythrocytes and platelets are two major MP replication populations in the circulation of DENV-infected Rearrangement of membrane structure induced by patients. Elevated levels of red blood cell-derived MPs dengue virus (DENV) is essential for replication, and (RMPs) directly correlated with DENV disease severity, requires host cellular machinery. Adaptor protein whereas a significant decrease in platelet-derived MPs complex (AP)-1 is a host component, which can be was associated with a bleeding tendency. Removal by Frecruited to components required for membrane mononuclear cells of complement-opsonized NS1- rearrangement. Therefore, dysfunction of AP-1 may anti-NS1 immune complexes bound to erythrocytes affect membrane organization, thereby decreasing via complement receptor type 1 triggered MP replication of virus in infected cells. shedding in vitro, a process that could explain the increased levels of RMPs in severe dengue. These In the study, it was demonstrated that AP-1-dependent findings point to the multiple roles of MPs in dengue traffic inhibitor inhibited DENV protein expression and pathogenesis. They offer a potential novel biomarker virion production. Researchers further clarified the candidate capable of differentiating dengue fever from role of AP-1A in the life cycle of DENV by RNA the more serious dengue hemorrhagic fever. interference. AP-1A was not involved in DENV entry into cells. However, it facilitated DENV RNA replication. This research was a collaborative work among RNAi specific to AP-1A decreased viral RNA and protein scientists from Songkhla Hospital, Khon Kaen Hospital, levels, and virion production in Huh7 cells. Huh7 cells Mahidol University and Medical Biotechnology transfected with AP-1A siRNA showed greater Research Laboratory. modification of membrane structures and fewer vesicular packets compared with cells transfected with Ref: Punyadee, N., Mairiang, D., Thiemmeca, S., Komoltri, C., Pan-Ngum, W., Chomanee, N., Charngkaew, K., Tangthawornchaikul, control siRNA. Therefore, AP-1A may partly control N., Limpitikul, W.,Vasanawathana, S., Malasit, P. and Avirutnan, P. DENV-induced rearrangement of membrane structures (2015). Microparticles provide a novel biomarker to predict severe required for viral replication. clinical outcomes of dengue virus infection. Journal of Virology, 89(3),1587-607.
This study was conducted by the Faculty of Medicine Siriraj Hospital, Mahidol University, National University of Singapore and Medical Biotechnology Research Laboratory. 22 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Highlights from Tuberculosis Rapid and low-cost detection platform for Research multidrug-resistant tuberculosis The traditional culture based drug susceptibility Genotypic diversity of drug-resistant tuberculosis testing (DST) is the primary diagnostic platform for isolates in Thailand multidrug-resistant tuberculosis (MDR-TB) in most Drug-resistant tuberculosis (TB), which includes developing countries. The consequent diagnostic multidrug-resistant (MDR-TB), quinolone-resistant time-delay is a major cause of escalating incidence. (QR-TB) and extensively drug-resistant tuberculosis The key to preventing further spread is early detection (XDR-TB), is a serious threat to TB control. To identify and treatment. While several molecular tests exist, whether the emergence of drug-resistant TB is they are limited by complexity and cost, hindering attributable to transmitted resistance or acquired their widespread application. resistance, researchers set to characterize the genotypic diversity of drug-resistant TB clinical isolates Researchers developed a simple Nucleic Acid Lateral collected in Thailand. Spoligotyping, a simple and Flow (NALF) immunoassay to complement rapid method for detecting polymorphisms within the conventional PCR, for the rapid molecular detection direct repeat locus, was used as a first-line genotyping of MDR-TB. The NALF device was designed using tool. A 24-locus MLVA was used for further subtyping antibodies for the indirect detection of labeled PCR of drug-resistant M. tuberculosis isolates. amplification products. Multiplex PCR was optimized to permit the simultaneous detection of the drug Results show the usefulness of genotypic diversity as resistant determining mutations in the 81-bp hot spot a supportive tool for understanding the causes of region of the rpoB gene (rifampicin resistance), while emergence drug-resistant TB over the study period. semi-nested PCR was optimized for the S315T Firstly, the clonal spread of MDR-TB and XDR-TB mutation detection in the katG gene (isoniazid provided the genetic evidence of previous MDR-TB resistance). This newly designed NALF is a simple, community outbreak and revealed a hidden spreading rapid and low-cost device suitable for low resource event of XDR-TB, respectively. Secondly, three major settings where conventional PCR is already employed clusters among all resistance groups suggested the on a regular basis. Moreover, the use of antibody- spread of predominant clones and amplification of based NALF to target primer-labels, without the resistance. Finally, a moderate number of unique requirement for DNA hybridization, renders the device genotypes hinted at the possibility of acquired generic, which could easily be adapted for the resistance as another factor driving the emergence molecular diagnosis of other infectious and non- of drug-resistant TB. The results of this study emphasize infectious diseases requiring nucleic acid detection. the need to strengthen TB control strategies and to adopt appropriate treatment regimens to prevent the further development of drug resistance.
This work was a collaborative study by researchers from the Faculty of Medicine Siriraj Hospital, Mahidol University, the Research Institute of Tuberculosis (Japan) and Medical Molecular Biology Research Unit.
Ref: Disratthakit, A., Meada, S., Prammananan, T., Thaipisuttikul, I., Doi, N. and Chaiprasert, A. (2015). Genotypic diversity of multidrug-, quinolone- and extensively drug-resistant Mycobacterium tuberculosis isolates in Thailand. Infection, Genetics and Evolution, 32, 432-9. Schematic illustration of Nucleic Acid Lateral Flow (NALF) design for the rapid detection of multidrug-resistant tuberculosis. Research and Development 23
This work was conducted by the Faculty of In this study, a modified continuous stirred tank Medicine Siriraj Hospital, Mahidol University, National reactor (CSTR) with a deflector installed at its upper Nanotechnology Center and Medical Molecular Biology section to promote the retention of suspended solids Research Unit. in the reactor was used to treat raw POME to produce biogas. Neither physical pretreatment nor chemical Ref: Kamphee, H., Chaiprasert, A., Prammananan, T., Wiriyachaiporn, N., pretreatment was performed to remove biofibers and Kanchanatavee, A. and Dharakul, T. (2015). Rapid Molecular Detection of Multidrug-Resistant Tuberculosis by PCR-Nucleic Acid FOG. This modified CSTR can be operated with raw Lateral Flow Immunoassay. PLOS ONE, 10(9): e0137791. doi:10.1371/ POME at the highest organic loading rate (OLR) of journal.pone.0137791. 19.0 g COD/l d, producing methane at 4.14 l/l d. Both cellulose and hemicelluloses were degraded at Highlights from Energy and significant rates in the modified CSTR. Environment This study was conducted by researchers from King Mongkut’s University of Technology North Bangkok, Modified reactor for treating raw palm oil mill King Mongkut’s University of Technology Thonburi effluent and Waste Utilization and Management Laboratory. The wastewater from palm oil production, commonly known as palm oil mill effluent (POME), typically Ref: Khemkhao, M., Techkarnjanaruk, S. and Phalakornkule, C. (2015). Simultaneous treatment of raw palm oil mill effluent and contains high levels of free fat, oil and grease (FOG), biodegradation of palm fiber in a high-rate CSTR. Bioresource and high concentrations of suspended solids and Technology, 177, 17-27. colloidal components, such as oil and biofibers, which pose problems in the operation of anaerobic processes Enhancement of starch-pulp separation in cassava for wastewater treatment and biogas production. starch production Either physical treatment or chemical treatment was In the cassava starch production process, starch employed to remove fine suspended solids and FOG granules following rasping step are divided into free from POME before processing in high-rate anaerobic and bound starch; the latter remains in the pulp and Freactors. However, it is desirable to investigate the is difficult to separate, while the former is not bound possibility of developing anaerobic reactors which can inside the pulp complex structure. In a starch extractor, process raw POME without chemical or physical cassava starch granules are separated from pulp pretreatments, to keep the process simple and reduce through the mechanisms of centrifugation and time, cost and the use of chemicals. filtration.
To obtain information on the starch granule characteristics and centrifugal-filtration process in cassava starch production, researchers investigated the effects of particle size and variety of cassava root, centrifugation and filtration mechanisms on free starch granule separation efficiency. Three cassava root varieties, Rayong 9, Rayong 11 and Kasetsart 50, were classified by particle size after grinding and sieving. Experiments were conducted at various relative centrifugal forces and pressure drops. The free starch separation efficiency increased with decreasing particle size of all cassava root varieties. The grinding of cassava root into small pieces caused cell wall breakage, facilitating free starch separation Scanning electron microscope photograph with 1000× magnification from the pulp. As the relative centrifugal force of cassava pulp, free starch granule, and bound starch granule. increased, some bound starch granules were released due to the force acting on the cassava pulp. The pressure drop in filtration process drove the free starch granules to pass through the screen although 24 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
this force was insufficient to separate the bound starch granules from the fiber. The results of this study will be useful for designing the improvement of starch extraction, thus promoting resource utilization efficiency in the starch factory.
This study was conducted by researchers from King Mongkut’s University of Technology Thonburi in collaboration with Waste Utilization and Management Laboratory.
Ref: Saengchan, K., Nopharatana, M., Lerdlattaporn, R. and Songkasiri, W. (2015). Enhancement of starch-pulp separation in centrifugal- filtration process: Effects of particle size and variety of cassava root on free starch granule separation.Food and Bioproducts Processing, 95, 208-217.
Highlights from Biodiversity Research and Utilization
Collection of biological materials BIOTEC established the BIOTEC Culture Collection (BCC) in 1996 to be a depositary and distribution center for microbes. The facility is also designated by the Department of Intellectual Property as a repository of patent-related microorganisms. At present, a total of 75,926 strains are preserved at the BCC, of which 63.44% are fungi, 26.77% are bacteria, 9.51% are yeast and 0.28% are algae. BCC on-line catalogue holds a total of 7,614 strains which are ready for distribution service to clients. All strains in BCC are maintained by freezing and/or freeze drying to ensure their genetic stability for long term storage. Precautionary steps have been taken to ensure the survival and integrity of the preserved cultures, including temperature control and regulator units, temperature monitoring with alert systems and back-up power-storage units. BCC has also been certified with ISO 9001:2008 as a service provider since 2005.
BIOTEC has also collected and preserved other biological materials under BIOTEC Molecular Genomic Materials Collection (BMGC) for both research and industrial utilization. A total of 216 biological materials are currently being preserved in the collection, comprising 45 vectors, 25 hosts and 146 recombinant clones. BIOTEC Bangkok Herbarium (BBH) preserves 39,860 dried specimens of fungi; whereas Hybridoma Bank has 644 monoclonal antibody-secreting Microbial Culture Stock Room for maintaining microbial strains in liquid nitrogen. hybridomas. Research and Development 25
In February 2015, BIOTEC officially launched Thailand • Six new yeast species: Yamadazyma insecticola f.a., Bioresource Research Center (TBRC) with the main sp. nov. (isolated from frass of an unidentified mission to promote accessibility and utilization of insect), Yamadazyma epiphylla f.a., sp. nov. (isolated biological materials, operating within a new concept from the external surfaces of rice leaves), Hannaella of a value co-creation and co-innovation approach siamensis sp. nov. (isolated from rice leaves), through network interaction. TBRC aims to hold Hannaella phetchabunensis sp. nov. (isolated from selected high quality biological materials with known corn leaf), Occultifur tropicalis f.a., sp. nov. (isolated characteristics for services to its clientele with an from sugarcane) and Hannaella phyllophila sp. nov. international quality standard “OECD best practice (isolated from plant leaves collected in Thailand and guidelines for biological resource center”. TBRC also Taiwan). forms partnerships with other culture collections Acetobacter locally and internationally, so that clients can access • Four new bacterial species: thailandicus and place an order for microorganisms from partner sp. nov. (isolated from a flower of the Sinosporangium fuscum collections through TBRC. At present, TBRC microbial blue trumpet vine), sp. catalogue contains 10,019 microbial strains from five nov. (isolated from an evergreen forest soil sample), Micromonospora fluostatini collections (TBRC, BCC, Thailand Institute of Scientific sp. nov. (isolated from Actinoplanes and Technological Research (TISTR), Chiang Mai near-shore sediment in Thailand) and luteus University and Khon Kaen University). These are sp. nov. (isolated from dry evergreen forest categorized as 2,143 strains of bacteria, 3,509 strains soil collected in Thailand). of filamentous fungi and 4,367 yeast strains, among these 472 are type strains. Other types of biological materials such as antibody and plasmid will be available for service in 2016. TBRC is also a member of the World Federation for Culture Collections (WFCC).
FMicrobial diversity study Microbial diversity study at BIOTEC involves the collection, isolation and identification of microorganisms from a variety of natural habitats. A total of fourteen novel species were discovered in 2015 and many of these discoveries were made in collaboration between researchers from BIOTEC and Two new species of Ophiocordyceps unilateralis: Ophiocordyceps other academic and research institutes. They are: septa (left) and Ophiocordyceps rami (right) on unidentified ants of the genus Camponotus. • Three new insect-pathogenic fungal species: Ophiocordyceps septa (isolated from unidentified ants of the genus Camponotus in Thailand), Ophiocordyceps rami (isolated from unidentified ants of the genus Camponotus in Thailand) and Aschersonia narathiwatensis (isolated from whitefly nymphs (Hemiptera) collected at Hala-Bala Wildlife Sanctuary in Thailand).
• A new genus and species of an aero-aquatic helicosporous fungus: Helicocentralis hyalina gen. et sp. nov.
A new species of insect-pathogenic fungi, Aschersonia narathiwatensis grown on whitefly nymphs (Hemiptera). 26 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Four new metabolites isolated from the Four new compounds isolated from the soil fungus seagrass-derived fungus Pestalotiopsis sp. PSU-ES194. Penicillium copticola PSU-RSPG138.
Bioactive compound discoveries • Two new hydroanthraquinones, paradictyoarthrins A and B, were isolated from the mangrove-derived BIOTEC natural product discovery program aims to fungus Paradictyoarthrinium diffractum BCC 8704. explore microbial resources for bioactive compounds These compounds exhibited cytotoxic activities. that have potential for pharmaceutical and agricultural product development. Bioactive substances produced • Four new metabolites, including two meroterpenoids, from various microorganisms are identified by activity- an isocoumarin, and a phenol, were isolated from guided fractionation and subjected to spectroscopic the seagrass-derived fungus Pestalotiopsis sp. PSU- analyses to determine chemical structures. In 2015, ES194 together with eight known compounds. The a total of 133 active compounds were identified, 40 biosynthetic pathway for isolated meroterpenoids of which were novel compounds. Below are examples is proposed and employed to establish their of microbial-derived compounds that have been absolute configurations. The isolated compounds identified in 2015: were evaluated for antimicrobial, antimycobacterial, antimalarial and cytotoxic activities. • Two new cyclic carbonate derivatives, aspergillusols A and B, one new eutypinic acid derivative, • One new hydronaphthalenone derivative was aspergillusic acid, two new phenalenones, isolated from the broth extract of the endophytic aspergillussanones A and B, one new cytochalasin, fungus Daldinia eschscholtzii PSU-STD57, together aspergilluchalasin, and one new γ-butyrolactone, with five known compounds. All the compounds aspergillulactone, along with six known secondary were tested for antimicrobial activity against metabolites were isolated from the soil Staphylococcus aureus, methicillin-resistant S. fungus Aspergillus sp. PSU-RSPG185. Aspergillusols aureus and Microsporum gypseum. A and B contain an unusual cyclic-carbonate • Six new compounds, an N-hydroxypyridone functionality; whereas aspergillussanones A glucoside, orbiocrellin A, its aglycone orbiocrellin B, exhibited weak activity toward KB and Vero cells. chromone glucosidesd, a dihydrochromone and a • Four new compounds including two eremophilane chromone, were isolated from the scale-insect sesquiterpenes, penicilleremophilanes A and B, as pathogenic fungus Orbiocrella sp. BCC 33248. well as two sulfur-containing biphenols, Orbiocrellin A exhibited antimalarial activity penicillithiophenols A and B, were isolated from the against Plasmodium falciparum K1 while it was non- soil fungus Penicillium copticola PSU-RSPG138 cytotoxic. In contrast, orbiocrellin B showed both together with 16 known compounds. Their structures antimalarial and cytotoxic activities. were elucidated by spectroscopic methods. Known • Two new ascochlorin derivatives, nectchlorins A and sporogen AO-1 and penicilleremophilanes A B, together with eight known compounds, were exhibited antimalarial activity against Plasmodium isolated from cultures of the leafhopper pathogen falciparum. Microcera sp. BCC 17074. Cytotoxic activities of these ascochlorin derivatives were evaluated. Research and Development 27
• Four new compounds, which were (−)-(7R)-7-O- methylsydonic acid, gibellulic acid, gibellulins A and B, together with 14 known compounds were isolated from the insect pathogenic fungus, Gibellula sp. BCC36964. These compounds were evaluated for the biological activities including anti-bacterial, anti- phytopathogenic, anti-Herpes simplex virus-type 1 activities, and cytotoxicity against both cancerous and non-cancerous cells.
• Four quinazolinones, new naturally occurring and three known derivatives, along with previously synthesized 4-phenylbut-3-enamide and three known compounds were isolated from an actinomycete, Streptomyces sp. BCC 21795. The new compound exhibited strong cytotoxic activity to Vero cells.
• Two new prenylhydroquinone-derived compounds, Ientinospirol and 1-(2,5-dihydroxyphenyl)-4- hydroxy-3-methyl-l-butanone, were isolated from cultures of the basidiomycete Lentinus similis BCC 52578, together with five known compounds.
• A new naphthoquinone, solaninaphthoquione, and a new succinate ester derivative, 4- Ophiocordyceps polyrhachis-furcata is a species in the O. unilateralis (4-hydroxyphenethoxy)-4-oxobutanoic acid, were species complex specific to the ant Polyrhachis furcata. F isolated from the soil fungus Fusarium solani PSU- RSPG227 together with five previously reported compounds.Solaninaphthoquione showed BIOTEC researchers reported a draft genome of O. significant cytotoxic activity against breast cancer polyrachis-furcata, a species in the O. unilateralis (MCF-7) cells, mild cytotoxic activity against oral species complex, which is also a Hypocrealean human carcinoma (KB) cells and weak antimalarial entomopathogenic fungus. Comparative analyses on activity. genes involved in pathogenicity and virulence between O. polyrhachis-furcata and other fungi were • Twelve aromadendrane sesquiterpenoids, inonotins performed. The research team identified genes A–L, and a previously unknown cyclofarnesane, i.e., involved in various steps of pathogenesis, investigated inonofarnesane, together with two known the common attributes of being entomopathogenic compounds, were isolated from cultures of the and the extent to which the host ranges have shaped wood-rotting basidiomycete Inonotus sp. BCC their evolution as well as enabling the discovery of 23706. new biosynthetic pathways. Insights to pathogenicity and host specificity in insect fungi This study was a collaborative effort between Bioresources Technology Research Unit and Genomic Ophiocordyceps unilateralis is referred to as a zombie Technology Research Unit. ant fungus because it causes the infected host ant to
climb into vegetation, bite vegetal materials then hang Ref: Wichadakul, D., Kobmoo, N., Ingsriswang, S., Tangphatsornruang, S., themselves upside down until death. The biology of O. Chantasingh, D., Luangsa-ard, J.J. and Eurwilaichitr, L. (2015). Insights unilateralis still has much to be discovered and the from the genome of Ophiocordyceps polyrhachis-furcata to pathogenicity and host specificity in insect fungi.BMC Genomics, 16, genome sequencing of this species will help gain an 881. insight. 28 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Microbial diversity in flood areas during Thailand’s Ref: Mhuantong, W., Wongwilaiwalin, S., Laothanachareon, T., Eurwilaichitr, L., Tangphatsornruang, S., Boonchayaanant, 2011 flood B., Limpiyakorn, T., Pattaragulwanit, K.,Punmatharith, T., McEvoy, J., Khan, E., Rachakornkij, M. and Champreda, V. (2015) Survey of In late 2011, Thailand experienced one of the most Microbial Diversity in Flood Areas during Thailand 2011 Flood Crisis damaging floods of the century. Flooding spread Using High-Throughput Tagged Amplicon Pyrosequencing. PLOS through the provinces of Northern, Northeastern and ONE, 10(5):e0128043. Central Thailand along the Mekong and Chao Phraya river basins and lasted for months in some areas, Study of polyketide synthase (PKS) genes in especially in the Central Plain. The flood ecosystem is entomopathogenic fungi considered a temporally variable ecological-niche Entomopathogenic fungi are able to invade and kill within the aquatic environment that can have a major insects. Various secondary metabolites can mediate impact on microbial communities related to water the interaction of a fungal pathogen with an insect quality and public health. host and also help the fungus compete with other microbes. In this study, bacterial and fungal diversity in sediments and waters collected from ten flood areas in Bangkok Researchers screened 23 isolates of entomopathogenic and its suburbs, covering residential and agricultural fungi for polyketide synthase (PKS) genes and areas, were analyzed using high-throughput 454 amplified 72 PKS gene fragments using degenerate pyrosequencing of 16S rRNA gene and internal PCR. A phylogenetic analysis was performed and 72 transcribed spacer sequences. Analysis of the microbial PKSs were identified from four insect fungal genome community showed differences in taxa distribution in sequences. The resulting genealogy indicated 47 water and sediment with variations in the diversity of orthologous groups with 99–100% bootstrap support, saprophytic microbes and sulfate/nitrate reducers suggesting shared biosynthesis of identical or closely among sampling locations, suggesting differences in related compounds from different fungi. Three insect- microbial activity in the habitats. Overall, specific groups were identified among the PKSs in Proteobacteria represented a major bacterial group reducing clades IIa, IIb, and III, which comprised PKSs in waters, while this group co-existed with Firmicutes, from 12, 9, and 30 fungal isolates, respectively. A IIa- Bacteroidetes, and Actinobacteria in sediments. IIb pair could be found in seven fungi. Expression Anaeromyxobacter, Steroidobacter, and Geobacter analyses revealed that eleven out of twelve PKS genes were the dominant bacterial genera in sediments, identified in Beauveria bassiana BCC 2660 were while Sulfuricurvum, Thiovirga, and Hydrogenophaga expressed in culture. PKS genes from insect-specific predominated in waters. For fungi in sediments, clades IIa and IIb were expressed only in insect- Ascomycota, Glomeromycota, and Basidiomycota, containing medium, while others were expressed only particularly in genera Philipsia, Rozella and in PDB or in CYB, PDB and SDY. The data suggest the Acaulospora, were most frequently detected. potential production of several polyketides in culture. Chytridiomycota and Ascomycota were the major As fungal polyketides from these entomopathogens fungal phyla, and Rhizophlyctis and Mortierella were represent valuable natural product resources, these the most frequently detected fungal genera in water. PKS genes’ phylogeny and expression data would be Diversity of sulfate-reducing bacteria, related to odor important for further studies of valuable polyketides problems, was further investigated using analysis of in the future. the dsrB gene which indicated the presence of sulfate- reducing bacteria of families Desulfobacteraceae, This study was performed by Bioresources Technology Desulfobulbaceae, Syntrobacteraceae and Research Unit in collaboration with King Mongkut’s Desulfoarculaceae in the flood sediments. The work University of Technology Thonburi and Assumption provides a foundation for more detailed studies on University. how microbial communities in flood ecosystems affect physicochemical changes in the environment and Ref: Punya, J., Swangmaneecharern, P., Pinsupa, S., Nitistaporn, P., Phonghanpot, S., Kunathigan, V., Cheevadhanarak, S., impact human communities. Tanticharoen, M. and Amnuaykanjanasin, A., (2015). Phylogeny of type I polyketide synthases (PKSs) in fungal entomopathogens and This study was the result of collaborative work by expression analysis of PKS genes in Beauveria bassiana BCC 2660. Fungal Biology, 119(6), 538–550. researchers from Chulalongkorn University, North Dakota State University (USA) and BIOTEC. Research and Development 29
Role of tenellin in iron homeostasis in Beauveria gene is a promising tool for further reconstitution of bassiana the fatty acid profile, in a host system of choice, for the production of economically important fatty acids, Iron is an essential element for life. However, iron particularly the n-3 and n-6 polyunsaturated fatty overload can be toxic. In this study, researchers acids. investigated the significant increase of tenellin and iron-tenellin complex production in ferricrocin- deficient mutants ofBeauveria bassiana. The chemical This work was conducted by Bioresources Technology analysis indicated that the ferricrocin-deficient Research Unit.
mutants nearly abolished ferricrocin production. In Ref: Jeennor, S., Cheawchanlertfa, P., Suttiwattanakul, S., Panchanawaporn, turn, these mutants had significant accumulation of S., Chutrakul, C. and Laoteng, K. (2015). The codon-optimized iron-tenellin complex in their mycelia under iron- Δ6 -desaturase gene of Pythium sp. as an empowering tool for engineering n3/n6 polyunsaturated fatty acid biosynthesis. BMC replete condition. Both tenellin and iron-tenellin Biotechnology, 15, 82. complex were not detected in the wild-type under such condition. Mass analysis of the mutants’ crude extracts demonstrated that tenellin formed a 3:1 Highlights from Biorefinery complex with iron in the absence of ferricrocin. The unexpected link between ferricrocin and tenellin Binding characteristic and synergistic effect of biosynthesis in ferricrocin-deficient mutants could be expasins on cellulose degradation a survival strategy during iron-mediated oxidative Degradation of lignocellulosic plant biomass is a key stress. bio-geochemical process in the organic carbon cycle. Expansins are non-hydrolytic proteins which have This study was conducted by Bioresources Technology been demonstrated to enhance the hydrolytic Research Unit and King Mongkut’s University of efficiency of lignocellulose by loosening the plant cell Technology Thonburi. wall structure. The molecular mechanism of how expansins enhance cellulose degradation is unclear Ref: Jirakkakul, J., Cheevadhanarak, S., Punya, J., Chutrakul, C., Senachak, F J., Buajarern, T., Tanticharoen, M. and Amnuaykanjanasin, A. (2015). and limited study has been performed on their Tenellin acts as an iron chelator to prevent iron-generated reactive substrate binding characteristics and interaction with oxygen species toxicity in the entomopathogenic fungus Beauveria bassiana. FEMS Microbiology Letters, 362(2), 1-8. cellulase-degrading enzymes.
Powerful tool for engineering polyunsaturated Researchers identified the binding characteristics of fatty acid biosynthesis five expansins originating from different groups of bacteria on cellulosic and hemicellulosic 6 ∆ -Desaturase is a key enzyme involved in the polysaccharides. The synergistic action of these biosynthesis of polyunsaturated fatty acids (PUFAs). expansins with Trichoderma reesei cellulase on To manipulate the oil composition of organisms of different types of polysaccharides was also studied. choice, by a metabolic engineering approach, a high The work provides an insight into the biological 6 expression level of the ∆ -desaturase enzyme in a functions of expansins on plant cell wall degradation heterologous host is required. As well as optimized with the potential uses on enhancing degradation and culture conditions, an efficient promoter and substrate modification of plant lignocellulosic polysaccharides availability, codon optimization is one of the most in bio-industries. common approaches for improving heterologous gene expression. This study was performed by Bioresources Technology Research Unit in collaboration with Kasetsart 6 Researchers cloned a ∆ -desaturase gene University. from Pythium sp. The gene encoded an enzyme which catalyzed the ∆6 -desaturation of the fatty acyl Ref: Bunterngsook, B., Eurwilaichitr, L., Thamchaipenet, A. and substrates having ∆9 -double bond. The product yields Champreda, V. (2015). Binding characteristics and synergistic effects of bacterial expansins on cellulosic and hemicellulosic substrates. were markedly enhanced by codon optimization of Bioresource Technology, 176, 129-135. the Pythium gene. This codon-optimized 6∆ -desaturase 30 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Ethyl acetate showed comparable efficiency in separation of biomass components compared with methyl isolbutyl ketone (MIBK) as used in the conventional process. In addition, microwave treatment was used to replace conventional heating, with the advantages of lower formation of inhibitory by-products. The work provides an efficient alternative approach for separation of primary lignocellulosic components with high recoverability and selectivity for further valorization in integrated biorefineries.
This study was performed by Bioresources Technology Research Unit, King Mongkut’s University of Technology Thonburi, PTT Research and Technology Institute and PTT Group Frontier Research Center.
Ref: Imman, S., Arnthong, J., Burapatana, V., Champreda, V. and Laosiripojana, N. (2015). Fractionation of rice straw by a single-step solvothermal process: Effects of solvents, acid promoters, and microwave treatment. Renewable Energy, 83, 663-673.
Highlights from Bioinformatics
SpirPro: Spirulina-Proteome Repository Cyanobacteria have been experimentally used as a model for plants and bacteria, therefore many cyanobacteria, such as Synechocystis sp., Synechococcus sp., and Spirulina (Arthrospira) platensis, have been studied at the genomic and proteomic levels.
In the quest to understand the effect of fluctuating environmental temperatures on Spirulina mass cultivation, researchers performed proteomic analyses of Spirulina under optimal and temperature stress conditions. The study led to the development of Scanning electron micrographs of native and solid residues from SpirPro, an integrated database for Spirulina. This fractionation of rice straw. (A) native rice straw; (B) conventional database provides possible mechanisms, in terms of heating process; (C) microwave heating process i n the absence of acid promoter. protein-protein interaction networks and proteome- wide expression levels, underlying the temperature stress response of this cyanobacterium for use as Modified Clean Fractionation method model mechanisms for other photosynthetic Fractionation of lignocellulosic constituents is a organisms, such as plants, algae and other prerequisite for efficient utilization of plant biomass in cyanobacteria. Moreover, proteome-wide domain integrated biorefineries. Several techniques have been identification is available in the database, which might developed for biomass fractionation, including a newly- be useful for further studies on protein-protein reported method termed Clean Fractionation (CF). interaction domain analyses. SpirPro is publicly available on-line at http://spirpro.sbi.kmutt.ac.th. In this study, researchers tested different organic solvents and acid promoters for their effects on This work was a joint study between Biochemical fractionation of rice straw using a modified CF process. Engineering and Pilot Plant Research and Development Research and Development 31
Laboratory and King Mongkut’s University of Technology Thonburi.
Ref: Senachak, J., Cheevadhanarak, S. and Hongsthong, A. (2015). SpirPro: A Spirulina proteome database and web-based tools for the analysis of protein-protein interactions at the metabolic level in Spirulina (Arthrospira) platensis C1. BMC Bioinformatics, 16:233.
Comprehensive database for human–mouse comparative study of microRNA–promoter interactions In 2012, microRNA (miRNA)–promoter interaction resource (microPIR) was introduced as a comprehensive public database containing over 15 million predicted miRNA target sites located within human promoter System overview of microPIR2 illustrating three major components: sequences. The database assists experimental MySQL databases, web interface and the output link-outs. scientists in exploring interactions of interest and facilitates research in this area. Since the release, access to the predicted targets in the database has markedly increased, demonstrating the potentially increasing interest in the research of this topic and Highlights from Diagnostic the potential value of the microPIR database in the Technology future. Bead array for Listeria detection Given the conservation of miRNAs and associated Bacteria of the genus Listeria currently contain six gene regulatory mechanisms across species, it is species, but only L. monocytogenes is pathogenic to plausible that the regulatory role of miRNA through humans, causing listeriosis. Well established traditional Fpromoter recognition is also present among methods are tedious and time consuming and thus mammalian species. Researchers therefore made are not suitable for screening large numbers of food modifications and updated the database, naming it samples required to meet increasing global demand. microPIR2, which contains predicted miRNA promoter Some high-throughput methods based on detecting targets in the mouse genome. microPIR2 provides nucleotide sequences need sophisticated detectors, approximately 80 million human and 40 million mouse elaborate sample extraction, and highly-trained predicted target sites. In addition to being a reference personnel. Therefore, immunoassays are still preferred database, microPIR2 is a tool for comparative analysis by many end-users for high volume sample screening. of target sites on the promoters of human–mouse orthologous genes. In particular, this new feature was Bead array for L. monocytogenes detection has been designed to identify potential miRNA–promoter developed using specific monoclonal antibodies. The interactions conserved between species that could novel highly specific antibodies were obtained from be stronger candidates for further experimental hybridoma libraries generated by using formalin-killed validation. Additional supporting information such as and heat-killed L. monocytogenes as immunogens. nuclear and cytoplasmic localization of miRNAs and The bead array was able to detect the bacteria with miRNA–disease association is also incorporated into the same accuracy as the standard plating method at this new database. the 1 CFU level after only 24 h of the enrichment period. In addition, Listeria-specific 3C3 and L. microPIR2 was developed by Biostatistics and monocytogenes-specific 7G4 antibodies were Bioinformatics Laboratory. The database is available successfully employed to construct a multiplex on this website: http://www4a.biotec.or.th/micropir2. detection forListeria , Salmonella and Campylobacter in a bead array format by combining with commercial Ref: Piriyapongsa,J., Bootchai, C., Ngamphiw, C. and Tongsima, S. (2014). Salmonella-specific and available Campylobacter- microPIR2: a comprehensive database for human–mouse comparative study of microRNA–promoter interactions. Database, Vol. 2014: specific antibodies. article ID bau115; doi:10.1093/database/bau115. 32 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
This study was a collaborative work between Biosensing Technology Research Unit, Animal Biotechnology Research Unit and Queen’s University Belfast (UK).
Ref: Karoonuthaisiri, N., Charlermroj, R., Teerapornpuntakit, J., Kumpoosiri, K., Himananto, O., Grant, I. R., Gajanandana O. and Elliott, C. T. (2015). Bead array forListeria monocytogenes detection using specific monoclonal antibodies. Food Control, 47, 462-471.
Bovine embryo sex determination by multiplex loop-mediated isothermal amplification The ability to determine the sex of cattle embryos Schematic illustration of multiplex LAMP for before transfer is useful for livestock management, bovine embryo sex determination. particularly in the dairy cattle industry where female calves are preferred. The current methods for sexing This study was conducted by Animal Biotechnology bovine embryos normally rely on detecting Y Research Unit, Food Biotechnology Research Unit, chromosome–specific DNA using PCR or fluorescence in Chiang Mai University and Suranaree University of situ hybridization (FISH). However, these techniques Technology. are not widely used in the field as they are time consuming and labor intensive and require expensive Ref: Khamlor, T., Pongpiachan, P., Parnpai, R., Punyawai, K., Sangsritavong, S. and Chokesajjawatee, N. (2015). Bovine embryo sex determination instruments. Alternatively, a loop-mediated isothermal by multiplex loop-mediated isothermal amplification.Theriogenology , amplification (LAMP) technique can be used. However, 83(5), 891-6. two separate LAMP reactions, one for the male- specific detection and the other for the control New method for white spot syndrome virus reaction, are required for complete bovine sexing. detection using graphene oxide Graphene oxide (GO) is attractive for biological or Researchers developed a modified LAMP in a multiplex medical applications due to its unique electrical, format (multiplex LAMP) for highly efficient bovine physical, optical and biological properties. In particular, embryo sexing. The protocol can simultaneously its fluorescence quenching property has been detect both male-specific DNA and control DNA in a exploited for biosensing via fluorescence resonance single reaction tube using multiplex LAMP. Two energy transfer (FRET) mechanism. chromosomal regions, one specific for males (Y chromosome, S4 region) and the other common to Researchers developed a white spot syndrome virus both males and females (1.715 satellite DNA), were (WSSV) DNA detection method based on loop- amplified in the same reaction tube. Each target was mediated isothermal amplification (LAMP) combined amplified by specifically designed inner primers, outer with FRET between GO and fluorescein isothiocyanate- primers and loop primers, where one of the S4 loop labeled probe (FITC-probe). The technique has a primers was labeled with the fluorescent dye detection limit of 10 copies of WSSV plasmid DNA or 6-carboxyl-X-rhodamine (emitting a red color), 0.6 fg of DNA extracted from shrimp infected with whereas both satellite loop primers were labeled with WSSV and no cross contamination. the fluorescent dye fluorescein isothiocyanate (emitting a green color). After amplification at 63°C The results demonstrated that GO-FRET with LAMP for 1 hour, the amplified products were precipitated technique is promising for fast, sensitive and specific by a small volume of cationic polymer predispensed DNA detection and could be applied to a point-of- inside the reaction tube cap. Green precipitate operating system. indicated the presence of only control DNA without the Y chromosome, whereas orange precipitate indicated the presence of both target DNAs, enabling interpretation as female and male, respectively. The multiplex LAMP showed 100% accuracy in identifying the actual sex of the embryos. The method is cost Schematic representation of LAMP-GO-FRET assay for fluorescence detection of white spot syndrome virus DNA. effective and highly mobile, making it suitable for field use. Research and Development 33
This study was conducted by Biosensing Technology for commercial purpose and has various industrial Research Unit and National Electronics and Computer applications in the value chain analysis, thus yield Technology Center. improvement through hybrid technology will help growers and industries gain substantial advantage. Ref: Waiwijit, U., Phokaratkul, D., Kampeera, J., Lomas, T., Wisitsoraat, A., Farmers will likely adopt the technology if the hybrid Kiatpathomchai, W. and Tuantranont, A. (2015). Graphene oxide based fluorescence resonance energy transfer and loop-mediated line performs far better than pure lines. To promote isothermal amplification for white spot syndrome virus detection. its technological capability in hybrid rice, an R&D and Journal of Biotechnology , 212, 44-49. capacity building plan needs to be in place, along with the plan to promote hybrid rice industry. Highlights from Policy Research The report of this study was submitted to Thailand Study on the potential of hybrid rice technology in Research Organizations Network, a network of seven Thailand government research funding agencies.
It is well documented that hybrid rice varieties yield Technology Action Plans for the agricultural sector to about 15 to 20 percent more than even the best of adapt to climate change the improved or high-yielding bred varieties do. Moreover, hybrid technology expedites breeding Agriculture remains a fundamental part of the Thai several desired traits into one variety. Hybrid rice economy and social structure. For Thailand, climate technology has been well exploited in China, and many change could therefore have significant effect on the rice-producing countries are now turning their agricultural sector, far more than any other sectors. attentions to hybrid rice study in recognition of its Three technologies have been identified as crucial good potential to improve food security and technologies in defining how Thailand can respond competitiveness. Hybrid rice however, is not well to, and mitigate the effects of, climate change on the adopted by farmers in Thailand because of the poor agricultural sector. They are: 1) forecasting and early performance of imported hybrid seeds under the local warning systems, 2) crop improvement and 3) Fclimate and conditions and the high price of domestic precision farming. Subsequently, Technology Action seed due to the lack of efficient hybrid seed production Plans (TAPs) were formulated for the next ten years, technology. Despite putting continuous effort into rice 2015-2025. TAPs comprise two sections: 1) Technology breeding, yield growth of Thai rice is declining, Action Plans for building research capability and indicating the limitation of pure line cultivars. The human resources, including priority crops and model question remains whether Thailand should pay more crops, for each technology, and 2) Action Plans for attention to this technology. Technology Transfer which address the cooperation among government agencies, academic sector, private BIOTEC research team conducted a study on the sector and farmers and the funding to support learning potential of hybrid rice technology in Thailand centers at the district level to deliver training modules focusing on three aspects: 1) market opportunity 2) for government officers (smart officers) and farmers farmers’ adoption and 3) technological capability. The (smart farmers) and manage knowledge; all of which study revealed that common white rice, among several go to ensure that the technologies are adopted and types of rice grown in Thailand, has potential for exploited. employing hybrid seed technology, as it is cultivated TAPs have been submitted to the National Science Technology and Innovation Policy Office, and have been incorporated into the Thailand Climate Change Master Plan 2015-2050 (led by Ministry of National Resources and Environment) and The Twelve National Agriculture and Cooperatives Plan 2017-2021 (led by Ministry of Agriculture and Cooperatives). Thailand Research Organizations Network, comprising seven government research funding agencies, has included climate change as a new target research initiative. Hybrid rice seed production fields at Pathum Thani Rice Research Center, a collaborative work between BIOTEC and the Rice Department. 34 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Technology Transfer 35
FTechnology Transfer
BIOTEC places strong emphasis on exploiting our research through the transfer of technologies to public or private sectors. The main mechanisms used to implement the technology transfer include technology and product licensing, capacity building, along with collaborative and commissioned research, as well as consultancy service. 36 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Licensing Agreements The licenses for the following technologies were extended during fiscal year 2015: The following technologies were licensed during fiscal year 2015: Air filter for tissue culture laboratory. An air filter for greenhouses or tissue culture containers, developed Production technology for low-cyanide cassava flour. by BIOTEC researchers, creates an environmental The technology is able to process bitter cassava, condition that allows for air and moisture exchange known to contain high levels of toxic cyanide, into in the tissue culture process, keeping the moisture high quality cassava flour with less than 10 ppm content within the tissue culture container at 45-65%, cyanide content. This technology can be installed in an optimal level for plant growth. The technology was the cassava starch factory with some process licensed to a tissue culture business operator in 2008 modification, enabling the factory to diversify its and the agreement was renewed with the same products. Chorchaiwat Industry Company Limited, a operator in 2015. starch-processing company, obtained the license to use this technology to produce cassava flour for Alpha thalassemia immunochromatographic strip market testing. test. The immunochromatographic strip is a qualitative, lateral flow immunoassay for the screening of various Colloidal solution as an indicator for acetone gas. types of alpha thalassemia traits by using whole blood. This technology involves two types of reagents which The test strip includes antibodies specific to the when mixed will react specifically with acetone gas gamma 4 protein and provides easy visual resulting in a color change. The detection range of discrimination between a positive result and a acetone is between 0 to 100 ppm. Health Innovision negative result. The technology was licensed to i+MED Company Limited has licensed the technology in order Laboratories Company Limited in 2011 and the to further develop it into a breath analyzer for non- agreement was renewed in 2015. invasive glucose monitoring.
High quality cassava flour, which contains no gluten, can be used as a Immunochromatographic strip test for alpha thalassemia detection. potential starch-based ingredient in many gluten-free diets for people who have celiac disease, or are gluten sensitive. Technology Transfer 37
Collaborative and Commissioned Center. Mitr Phol Group is Thailand's and Asia's biggest sugar and bio-energy producer. The research Research partnership brings in complimentary expertise from In fiscal year 2015, BIOTEC conducted 72 projects in each party, namely BIOTEC’s advanced omic the form of collaborative and commissioned research, technologies and Mitr Phol’s competence in sugarcane as well as consultancy services. Of these, 29 projects breeding, to develop sugarcane elite lines with high were initiated in 2015. Collaboration and services sugar content, high yield, drought tolerance, longer covered a range of topics, such as screening for ratooning and good genetic background. The results microorganisms and enzymes for specific purposes of this project will enable the sugar industry to and the development of an industrial-scale production overcome shortage of supply and inconsistency of system, designing fermentation processes for food sugar content from different cane varieties and and feed products, breeding of commercial crops, locations throughout the year. improving wastewater treatment/biogas system, recovering value-added products from waste streams Screening for potential microorganisms for animal and diagnostic development for environmental and feed probiotics for Asia Star Trade. Asia Star Trade is agricultural applications. Following are some of the a local company with core business in animal and highlights: aquaculture feed production. The company commissioned BIOTEC to find microorganisms with Establishment of standard protocol for disease- probiotic potential in order to develop these into a resistance screening in tomato with the Asia & Pacific commercial feed supplement. Asia Star Trade already Seed Association.The Asia & Pacific Seed Association has a few feed enzyme products based on technologies (APSA) is the largest regional seed association in the developed by BIOTEC. Fworld, whose aim is to promote quality seed production and marketing in the Asia and Pacific Region. BIOTEC Developing formulations for commercial biocontrol collaborates with APSA to develop an efficient products of Beauveria bassiana for Ladda. Ladda is inoculation protocol for screening Tomato necrotic a leading agro-chemical company in Thailand. The ringspot virus (TNRV) and Capsicum chlorosis virus company commissioned BIOTEC to develop (CaCV), two major tospovirus species causing severe formulations for commercial biocontrol products damage to tomato plantations. The developed based on one Beauveria bassiana strain screened by protocol will greatly enhance the development of BIOTEC, which has been shown to exhibit efficacy disease-resistant tomato lines in the seed industry. against insect pests including green peach aphid (Myzus persicae), pink mealybug (Phenacoccus Integrative sugarcane breeding to increase sugar manihoti) and brown planthopper (Nilaparvata content with Mitr Phol Innovation and Research lugens).
Tomato necrotic ringspot virus (TNRV) and Capsicum chlorosis virus (CaCV) are the major tospovirus species detected in tomato. 38 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Open Lab for Industry Production technology for high- quality Beauveria for Thai farmers Open Lab is an activity to promote collaboration between laboratories and industries by inviting To utilize Thailand natural resources for pest control, companies to meet with BIOTEC scientists, learn about BIOTEC research team explores insect pathogenic BIOTEC expertise and the potential of biotechnology fungi, which are natural enemies of pest insects, to for improving industrial processes and products. Open discover potential strains that could become effective Lab often leads to collaborative research, commissioned biological control agents that pose minimal threat to research, or the provision of analytical services. In non-target organisms. From our studies, Beauveria 2015, a total of 6 open lab events were organized. bassiana BCC2660 has been shown to exhibit efficacy Held during the NSTDA Annual Conference (NAC), against insect pests including green peach aphid NSTDA Open House offered opportunities for the (Myzus persicae), mealybug (Pseudococcus cryptus) private sector to visit various laboratories located in and brown planthopper (Nilaparvata lugens). Thailand Science Park, including BIOTEC laboratories. Researchers then developed a solid-state fermentation Over two hundred companies -- in various industries process using rice grain as a substrate for the effective such as food and health food, agriculture, IT, chemicals production of Beauveria inoculum at low cost. The and energy -- participated in NSTDA Open House. In process and conditions have been optimized to yield addition, throughout the year, two open lab events maximum spore production. were dedicated to agriculture and food industry with a total of 40 companies participating. One event was Realizing that one of the tasks undertaken by the designed for 10 aquaculture companies with the Department of Agricultural Extension (DOAE) is to showcase of various technologies to support supply Beauveria inoculum to farmers to promote the aquaculture industry such as diagnostic technology use of biocontrol agents, NSTDA is collaborating with for aquatic animals and recirculating aquaculture DOAE aiming to build up the capacity in Beauveria system. production in the Department, as well as with farmers. This will be achieved by the transfer of technology developed and the effective strain (B. bassiana BCC2660) screened by BIOTEC to regional centers of DOAE responsible for inoculum production and distribution, as well as the training of farmers on Beauveria production from inoculum. The pilot project will focus on upgrading production facilities and training of staff of DOAE regional centers in Chiang Mai, Phitsanulok, Phrae and Nakhon Sawan provinces, as well as farmers in those respective areas. It is expected that these four sites will further transfer the technology to other centers to cover the whole country.
The pilot project was launched in Chiang Mai on 22 June 2015, with a meeting among staff from NSTDA, BIOTEC and DOAE to formulate a detailed action plan. In addition, a training workshop on Beauveria Open lab and business matching are among mechanisms used to inoculum production was held for DOAE staff on the promote technology transfer and external collaboration. same day. Technology Transfer 39
Transfer of starter culture technology for silage production for cattle farms
In response to increasing demand for animal feed in the livestock sector, significant progress has been made in the improvement of forage crops in Thailand. Fast-growing and high-yield varieties of forage cane and napier grass have already been developed by BIOTEC-Kasetsart University and the Department of Livestock Development, respectively. However, feed shortage still persists during the dry season. To solve this problem, researchers from BIOTEC, Khon Kaen University and the Department of Livestock Development co-developed starter culture technology for ensilaging green fodder. Fast-growing lactic acid bacteria (LAB) were screened locally from the natural environment and used as starter culture to ensilage forage cane and napier grass when forages are high in quantity. It was found that, not only could the starter culture speed up the fermentation process, the silage Fproduced from starter culture technology is of higher quality and can be preserved for a longer period of time compared to naturally-fermented silage. This technology thus enables farmers to stockpile high- quality feed into the dry season.
Workshop on starter culture for silage production held for cattle This technology has been transferred to livestock farmers at the 2015 National Dairy Fair. farmers and feed producers at the 2015 National Dairy Fair. The workshop was also organized in Chiang Mai province to transfer the technology to farmers and operators in the northern region.
blueAmp, a simple and rapid detection test for streptococcosis in Nile tilapia, was introduced to potential investors at NSTDA Investors’ Day 2015. 40 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Human Resources Development 41
HUMAN RESOURCES FDEVELOPMENT
BIOTEC places a high priority on capacity building through increasing the quantity and quality of human resources in biotechnology as well as upgrading and educating the workforce. Several activities were designed to assist different segments of the workforce, as well as address a variety of objectives, ranging from providing fellowships, to training post-graduate students, to organizing scientific conferences and training workshops for academics and industry, as well as organizing youth programs. 42 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
International Conference on Anaerobic Digestion: AD Technology and Microbial Ecology for Sustainable Development (ADTech 2015) was held in Chiang Mai, Thailand.
In 2015, BIOTEC granted 11 post-doctoral fellowships, 3 of which are new fellowships. A total of 21 scientific events, including one international conference and twenty training workshops and seminars, were organized on various topics, such as plant breeding, microbial management, animal vaccine technology and guidelines for biosafety of genetically modified microorganisms. Science education was also promoted through the Science in Rural Schools (SiRS) program. In addition, BIOTEC also runs a program to build up capacity for farmers and community enterprises, offering training and technical assistance in Good Agricultural Practice and Good Hygiene Practice. Human Resources Development 43
International Conference on Foundation, the Netherlands), Prof. Théodore Bouchez (National Research Institute of Science and Technology Anaerobic Digestion for Environment and Agriculture, France), Prof. habil. The International Conference on Anaerobic Digestion: Bernd Bilitewski (INTECUS GmbH Waste Management AD Technology and Microbial Ecology for Sustainable and Environment-Integrating Management, Germany), FDevelopment (ADTech 2015) was held on 3-6 February Prof. Makarand Madhao Ghangrekar (Indian Institute 2015 in Chiang Mai, Thailand. The Conference was of Technology Kharagpur, India), Prof. Saulius organized by a joint effort of the Institut National de Vasarevicius (Vilnius Gediminas Technical University, la Recherche Agronomique (INRA), France, Chiang Mai Lithuania) and Prof. Morakot Tanticharoen (King University, King Mongkut’s University of Technology Mongkut’s University of Technology Thonburi, Thonburi and BIOTEC. Thailand).
ADTech 2015 provided a primary platform for in- depth Poster awards were announced on the last day of the presentation and discussion on various topics that Conference. The awards went to Prof. Kazuaki cover all aspects of anaerobic digestion namely Syutsubo (National Institute for Environmental microbial ecology in anaerobic environment; process Studies, Japan), Mr. Peerawat Khongkliang (Taksin engineering in anaerobic digestion; new biomass University, Thailand) and Asst. Prof. Piyanuch Niamsup feedstock for renewable energy; methane emission (Maejo University, Thailand) for gold, silver and bronze and climate change; biogas utilization and upgrading; prizes, respectively. and policy-economic aspects of anaerobic digestion. The Conference program consisted of 58 lectures and A visit to the Landfill Gas Energy Site was offered to 26 poster presentations. Keynote lecturers included participants as part of the excursion. ADTech 2015 Dr. Lodewijk Willem Hulshoff Pol (Lettinga Associates was well attended by 120 participants from 16 countries. 44 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Two training modules have been delivered in fiscal year 2015 while the rest will be offered in 2016.
Capacity Building in Biosafety of Genetically Modified Microorganisms
A genetically modified microorganism (GMM) is a microorganism whose genetic material has been altered using genetic modification techniques to Workshop participant studying root phenotypes produce desired genetic features for specific needs. in one of the plant breeding workshops. In an industrial scale, GMMs have been utilized to produce specially designed products in a wide range Building Up National Capacity in of industries, from pharmaceutical and medical supply industry, food industry, to bio-industry. To ensure the Plant Breeding human and environmental safety, the Technical Biosafety Committee (TBC) first released “Biosafety It has been reported that climate change will have a Guidelines for Contained Use of Genetically Modified great effect on agricultural sector. As Thailand is an Microorganisms at Pilot and Industrial Scales” back in agriculture-based economy, with half of the land and 2004, with regular revisions for the most up-to-date 35% of labor force devoted to agriculture, this is a information. The Guidelines cover the use of GMMs major threat to its economy and food security. To cope in containment at pilot and industrial scales according with the effect of climate change on food security, to GMM classification, together with suggested three technologies have been identified as crucial containment levels, GMM waste management, technologies for the development in the next 10 years: transport, possession, emergency plans and the plant breeding; precision farming; and forecasting and responsibilities of personnel associated with GMM warning systems. work. BIOTEC and NSTDA, in collaboration with several Thai Since the use of GMM has been on the rise, the universities, have developed an intensive capacity understanding and awareness among practitioners is building program in plant breeding in order to enhance necessary for the proper handling. BIOTEC, in human resource development in this particular area. collaboration with Faculty of Science, Chulalongkorn The Program consists of 6 training modules to be University and Faculty of Science, Mahidol University, offered through a series of workshops, namely: organized the Law and Regulation Workshop on How to Use Genetically Modified Microorganisms (GMMs) • Module I: Breeding of Cucurbit in Thailand on July 9, 2015, specifically targeting at • Module II: Technology Tools for Breeding: Corn regulatory officers and staff in the industry working Model with GMM. The Workshop was attended by 64 • Module III: Learning from Success Case of Rice participants from 16 governmental organizations and Marker-assisted Selection Breeding 11 companies in Thailand. • Module IV: Learning from Success Case of Fruit Tree • Module V: Plant Genomics / Molecular Tools • Module VI: Crop Breeding Concept and Success Case Human Resources Development 45
Training of Science Teachers in the Rural Area
BIOTEC initiated the Science in Rural Schools (SiRS) program in 1998. The establishment followed the initiative of HRH Princess Maha Chakri Siridhorn to improve the quality of life and enhance learning capability in science for rural students and science teachers, through activities such as producing educational media such as books, a website and CD- ROM, as well as supporting schools to organize science camps and science project contests to stimulate science learning.
Since 2009, SiRS has taken part in a larger initiative, the Little Scientists’ House Thailand, aiming to promote science education among children by offering training and learning materials to science educators and creating a community of practice among science teachers at the provincial, regional and national levels through the collaboration with the Haus der kleinen Forscher Foundation of Germany. SiRS has been Ftasked to disseminate the learning materials developed by Little Scientists’ House Thailand to schools in its network in four provinces (Maehongson, Sakon Nakon, Pangnga and Narathiwat), by developing training modules for teachers in rural schools to use the learning materials to fit with the context of the area and providing training to school teachers on how to use the learning media in the classroom. Workshops and follow-up meetings were organized with the pilot schools to monitor the improvement in students’ learning abilities. In 2015, 109 teachers participated in the training program. Training of science teachers to use learning materials in the classroom. 46 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Public Awareness 47
FPUBLIC AWARENESS
One of BIOTEC’s missions is to raise public awareness of how biotechnology and the life sciences relate to everyday life. This is part of the larger goal of making Thailand a knowledge-based economy. The mission is carried out in part by channeling information through popular media such as the internet and television, as well as providing information to journalists. BIOTEC also organizes exhibitions at various events, ranging from scientific conferences, as well as science and technology, agricultural and industry fairs. Lab tours are also organized to bring the public into BIOTEC research facilities where they can meet and talk to scientific staff. Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
ele s on Pro ra s
In BIOTEC rod ced one tele i ion rogra a in te ot for reg lar roadca t on lic channel a art of N T T total of ho ere rod ced and ent on air a ell a eing ade a aila le for ie ing on BIOTEC o T e (h yo t e co iotecthailand) The e tele i ion rogra er e e eral r o e fro rai ing a arene of cience ed cating the lic a o t the ene t of technologie de elo ed locally ho ca ing cce f l in ention that ha e reached the ar et and introd cing rod ct and technologie to otential in e tor or colla orator
Press cti ties
E ent ha e een organi ed reg larly for local o rnali t to e infor ed of o r cienti c re earch and the e of o r technologie in act al ind trie In e er of the re ere in ited to i it
Thana ai al a te tile factory that e EN ea e a d o acti ity en y e for one te iode i ing and io co ring roce of co on fa ric ai h nnatha a gra e rod cer that e N o rnali t i ited a ineyard ing N rod ct to control in ect e t to control in ect e t in the ineyard Organic rice far in a othorn ro ince here BIOTEC ro ote and de on trate the e of technologie to grade far ing ractice
Ne and article on the e torie ere e ently li hed reaching o t to the general lic
n article on ecology re earch a thored y BIOTEC re earch tea li hed in Bang o o t Public Awareness 49
BIOTEC exhibition at Thailand Lab 2015 in September.
Exhibitions Open Lab
BIOTEC participated in a number of exhibitions and Thailand Science Park, home of BIOTEC and its five displays at major events, disseminating results and research units, is considered the largest R&D ideas to several demographics and industrial sectors. community in Thailand, with government-funded Examples of key events were the 2015 National Dairy research labs and also private labs of several Day in January; IP Fair 2015 in February; NSTDA Annual companies. With such a concentration of facilities, FConference in April; Thai Rice Convention 2015 in May; Thailand Science Park makes an ideal study site for and Thailand Lab 2015 in September. schools, universities, government and private organizations, as well as the general public. BIOTEC has an open-door policy and welcomes the public to tour laboratory facilities on a regular basis.
In 2015, 124 groups toured BIOTEC laboratories. Visitors included faculty members and students from various universities in Thailand; participants from the Thailand-UK Workshop on Plant-microbe Interactions; executives of Thailand Rice Department; and representatives of Japan External Trade Organization (JETRO) and Japanese Chamber of Commerce in Thailand. 50 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) International Collaboration 51
INTERNATIONAL FCOLLABORATION
The BIOTEC International Cooperation Program aims to capitalize on international links to help BIOTEC and Thailand become a regional leader in the field of biotechnology. In so doing, the Center has developed close relations with overseas organizations at the bilateral, multilateral and regional levels. These relations are developed through formal collaborative agreements, organizing joint scientific seminars with international partners, hosting foreign scientists and students in laboratories, and organizing an annual meeting of the BIOTEC International Advisory Board. 52 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
BIOTEC and Thermo Fisher Scientific embark on the study of natural products for agricultural application.
Fostering Collaboration
In fiscal year 2015, BIOTEC signed seven new Memorandums of Understanding (MOUs)/agreements and renewed one MOU to foster collaboration with the following organizations:
Organization Detail Okinawa National College of Technology, To develop collaboration on cassava and starch technology. Japan Institut Pastuer, France To study a vaccine candidate for porcine epidemic diarrhoea virus. Earlham College, USA To support the placement of students from Earlham College in BIOTEC laboratories Institute of Microbiology, Chinese Academy To promote cooperation in the field of microbial biotechnology. of Sciences, China Food Industry Research and Development To foster cooperation in the field of microbial biotechnology. Institute, Taiwan Universiti Putra Malaysia, Malaysia To promote cooperation in the field of microbial biotechnology. Thermo Fisher Scientific Private Limited To establish metabolomic and dereplication strategies for natural product research Meijo University, Japan To support research collaboration on salt-tolerant rice. International Collaboration 53
Technical Assistance for the Thai-Lao Collaboration on Insect- Establishment of Culture pathogenic Fungi Collection Center in Nepal In November 2014, an MOU on Thailand-Lao The advance in biotechnological and bioscience Cooperation on Science and Technology was entered research enables the discovery of microbial utilization, into, with biotechnology listed among 14 fields of especially in essential industries such as cooperation between the two nations. BIOTEC and its pharmaceuticals, energy and agriculture. Therefore, Lao counterpart, Biotechnology and Ecology Institute increasingly, culture collection becomes an important (BEI), have agreed to collaborate on research in infrastructure for a bio-economy. biodiversity and ecology of insect-pathogenic fungi.
Nepal Academy of Science and Technology (NAST) is Under the collaborative framework, BIOTEC and BEI keen to establish a culture collection center as a part co-organized a Training Workshop on Collection, of their National Biotechnology Center. NAST Identification and Preservation of Insect-pathogenic recognized the expertise of BIOTEC in the management Fungi in Vientiane on 2-5 June 2015. The Workshop of microbial collections and thus contacted BIOTEC to provided both fundamental knowledge and hands-on seek collaboration on this topic. BIOTEC invited a practice into the study of insect-pathogenic fungi. The scientific team from NAST, led by Dr. Buddhi Ratna Workshop was well attended by 15 research staff from Khadge, NAST Secretary, to visit Thailand on 24-26 BEI, National University of Laos and Plant Protection August 2015 in order to learn first-hand of BIOTEC’s Center. experience in establishing and managing its culture Fcollection. The study visit focused on the management of microbial culture collection which includes activities such as the preservation, identification, quality control, data management, as well as legal management. NAST also had an opportunity to visit Thailand Institute of Scientific and Technological Research (TISTR) and KEEEN, a company specializing in bioremediation technology, to broaden their perspective on microbial collection and utilization.
BIOTEC will provide training to NAST research staff on culture collection management in 2016. Workshop on Insect Pathogenic Fungi in Lao PDR. 54 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Thailand-China Silk Road Cooperation
BIOTEC and the Institute of Microbiology, Chinese Academy of Sciences (IMCAS) jointly organized a Seminar on Thailand-China Silk Road Cooperation on 19 May 2015, with 70 participants from research and academic organizations in Thailand. The seminar introduced the Chinese research and management of microorganisms and provided an opportunity for Thai and Chinese researchers to explore collaboration. The Chinese delegation was led by Prof. Shuang-Jiang Liu,
An MOU between BIOTEC and the Institute of Microbiology, Chinese Director-General of IMCAS. Topics of presentation Academy of Sciences signifies the collaboration in microbiology field. included the CAS international cooperation, cooperation on microorganisms, China General Microbiology Culture Collection (CGMCC), WDCM global collaboration and yeast research.
On this occasion, BIOTEC entered into an MOU with IMCAS to promote research and capacity building in microbiology, especially on culture collection management, yeast, lactic bacteria, fungi and big data management of microorganisms.
Visit of Canadian Biopesticide Expert
BIOTEC invited Prof. Alan Watson, Director, Biopesticide Research Laboratory at McGill University, Canada to provide advice to BIOTEC’s biocontrol research and technology transfer program. His visit included a meeting with BIOTEC research team working on Beauveria bassiana and vegetative insecticidal proteins (Vips), inspection of project sites in Ayutthaya and Phrae provinces, to which the cultivation and spore production technology ofB. bassiana developed by BIOTEC has been transferred, and a private company keen to develop a commercial Beauveria biopesticide. Prof. Alan Watson and BIOTEC research team during a visit to a company. International Collaboration 55
International Exchange Programs
A number of programs have been established to host foreign researchers and students in BIOTEC laboratories. These are:
• Human Resource Development Program in Biotechnology for Asia Pacific. Operating since 2001, this program provides fellowships to young researchers from developing countries in the Asia- Pacific region to undergo research-based training in BIOTEC laboratories for a period of 3-6 months. Approximately 10-15 fellowships are granted Students from Soon Chun Hyang University attending annually. the training program at BIOTEC in 2015. • TWAS-UNESCO Associateship Scheme. Under this scheme, BIOTEC becomes one of 100 Centers of Excellence in the South to offer fellowships to foreign researchers from the South for training at, and fostering collaboration with, BIOTEC.
• International Student Internship Program. BIOTEC offers internship placements in laboratories for F undergraduate and graduate students from overseas. This includes students sent from our partner institutes, under an established agreement, as well as students applying directly to the program. Current partner institutes include Atma Jaya Catholic University, Temasek Polytechnic, National Taiwan University, Nanyang Polytechnic, University of Kent, Soon Chun Hyang University and City University of Hong Kong. Earlham College joined this Program in 2015.
In 2015, BIOTEC hosted a total of 15 researchers and 58 students from 15 countries (Vietnam, Indonesia, Malaysia, Singapore, India, Taiwan, China, Korea, Japan, Germany, the Netherlands, the UK, the US, Canada, Mexico). Among the 73 exchange researchers/ students, 8 visitors were on-going from the previous year. 56 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Impact of BIOTEC’s Output 57
IMPACT OF FBIOTEC’S OUTPUT
Every year, a number of projects are selected for detailed impact study. These are projects that have created technologies or products that have actually reached the end users, through various forms of technology transfer such as technology licensing, provision of consultancy and training and establishing core infrastructure that would advance research in academic and industrial communities. Impact is measured in terms income generated by our clients from products and technologies and, where appropriate, assessed for the impact to the nation’s socio-economy in the forms of import substitution, investment and employment generation. 58 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
In 2015, 49 projects were selected for impact Animal and aquaculture production. Technologies assessment. An estimated total of 5.27 billion Baht and products under the assessment include steroid- was generated from these projects. This amount was based synchronization of ovulation protocol for categorized as investment generation (127 million artificial insemination of cattle, shrimp breeding, a Baht), revenue generation to licensees or users (4.54 closed recirculating aquaculture system integrated billion Baht), cost reductions (509 million Baht) and with hybrid nitrification biofilter tanks and patented import replacement (91 million Baht). Tubular Denitrification Reactor (TDNR) and shrimp disease detection kits. These technologies have generated an impact of 350 million Baht.
Agriculture and Food
A total of 38 projects were evaluated, generating 1.54 billion Baht impact.
New rice varieties. This includes varieties that have been developed by the Rice Gene Discovery Laboratory, in collaboration with the Rice Department, namely blast-resistant glutinous rice (Thanyasirin) and flood- tolerant rice (Homcholasit). The two varieties were introduced to famers in four regions of Thailand
through the transfer of seed production technology. Recirculation aquaculture system equipped with a nitrification biofilter A total of 72 million Baht in income increment was and patented tubular denitrification reactor at a tilapia hatchery facility estimated. of a company.
Technologies for crop production. BIOTEC engaged in various projects on plant improvement and technologies for crop production such as sugarcane breeding, increasing the production of disease-free sugarcane, orchid tissue culture production, rooftop and vertical plant cultivation system, environmentally- controlled greenhouse design, reagents for plant disease detection and the production of an NPV biopesticide product. These technologies have generated additional income to farmers and industries at the value of 232 million Baht.
Training and services on hormone-based technology for ovulation synchronization and artificial insemination were provided to both large and small scale dairy farmers. The technology helps induce pregnancy in cows with reproductive system difficulties, thus increasing the value of each cow. Impact of BIOTEC’s Output 59
Food and animal feed. Technologies that have been used at the commercial scale include starter culture for traditional fermented pork sausage, production technology of fermented soybean meal for animal feed, production of pentosanase for animal feed, industrial-scale production of pickle mustard greens using starter culture. These technologies have created income to the companies and investment to the economy at the value of 224 million Baht.
Community development. BIOTEC works with partners and local authorities to upgrade farming and cottage industry in the rural communities using area- based approach. Appropriate technologies, mainly related to farming and food processing, are made Animal feed, comprising soybean meal fermented with Bacillus, based accessible to villagers with proper training. Examples on the technology licensed from BIOTEC-NSTDA. of projects include promotion of organic rice production in Yasothorn province, agroforestry farming in Phrae province and Good Hygiene Practice (GHP) training for community food processing industries. These projects have improved the farming practice and food products, resulting in better quality Fproduce and products. The impact from investment and additional income is estimated at 218 million Baht.
Remediation of saline land. The project was implemented in collaboration with SCG, Pimai Salt Co. Ltd., and various partners, using salt-tolerant varieties of tropical plants, as well as some economic crops screened and developed from BIOTEC laboratory, along with soil treatment with organics and minerals The training on Good Hygiene Practice (GHP) enables community to treat saline land in four provinces - Sakon Nakhon, enterprises to obtain food safety certification from FDA. Udon Thani, Nakhon Ratchasima and Khon Kaen. The project was able to turn saline land into arable land for rice and other crops. Economic impact totaled 444 million Baht. 60 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Health and Medicine
One project was evaluated and created an impact of 67 million Baht.
Vaccine development. Chimeric dengue virus strains developed by BIOTEC and partner universities were licensed to a company for further investigation and development into a commercial vaccine. This project has generated investment to the economy of 67 million Baht.
Environment
Seven projects were evaluated, resulting in an impact of 503 million Baht.
Efficiency improvement in cassava starch processing. Technology has been developed to improve the production efficiency in terms of energy and water consumption and raw material utilization in the cassava starch production process. The technology was introduced to cassava starch factories through practical training and advice to practitioners working in the factories. A total of 389 million Baht in savings was estimated from cost reduction and improved production yields.
Biogas production from agro-industrial waste. Wastewater treatment and biogas production technology was developed by EcoWaste, a joint lab between BIOTEC and King Mongkut’s University of Technology Thonburi. The technology has been implemented in agro-industry factories (cassava starch, palm oil mill, food processing), enabling wastewater compliance and energy cost savings of 78 million Baht.
Environmentally-friendly products. BIOTEC and a private company collaborated on a joint research project to develop a commercial bioremediation product based on oil-degrading microbes. The technology was subsequently licensed to the company for commercialization. Impact assessment was made Engineers gathered samples from the starch production line to on the revenue and the value of import substitution, determine and monitor resource and energy efficiency in a cassava starch factory. totaling 36 million Baht. Impact of BIOTEC’s Output 61
Infrastructure Establishment
Three projects were evaluated resulting in an impact of 3.16 billion Baht.
Establishment of proteomics facility. This facility was established at Thailand Science Park, providing support on proteomics to researchers on a collaborative basis. The savings from seeking such a service from overseas were estimated to 82 million Baht.
BIOTEC Proteomics Facility is fully equipped with 2-D gelelectrophoresis, Biopharmaceutical facility. This facility was established image processing and analysis and mass spectrometry. through collaboration between BIOTEC and King Mongkut’s University of Technology Thonburi (KMUTT) and is located at Bangkhuntien Campus of KMUTT. It offers contract manufacturing services for the manufacture of investigational drugs, vaccines and biopharmaceuticals for clinical research, as well as services on process optimization, scale up, fermentation processing development and downstream processing development. In this initial stage, the impact was assessed on the investment Ffrom private sector at 1 million Baht.
DNATEC Laboratory. The laboratory was jointly set up by BIOTEC and Kasetsart University, providing services in DNA fingerprinting to authenticate plant varieties. The authentication of jasmine rice and other commercial rice varieties facilitates the sale of rice and enables high quality varieties to command premium price. Impact of 3.07 billion Baht was estimated.
National Biopharmaceutical Facility located at King Mongkut’s University of Technology Thonburi is fully equipped for contract manufacturing of investigational drugs, vaccines and biopharmaceuticals for clinical research. 62 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC) Appendices 63
FAPPENDICES
List of Publications List of Intellectual Properties Honors and Awards Executives and Management Team 64 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
List of Publications
1. Ajambang, W., Ardie, S.W., Volkaert, H., Ngando- 9. Arunpanichlert, J., Rukachaisirikul, V., Phongpaichit, Ebongue, G.F. and Sudarsono, S. (2015). Comparative S., Supaphon, O. and Sakayaroj, J. (2015). Xylariphilone: expression profiling of three early inflorescence stages a new azaphilone derivative from the seagrass-derived of oil palm indicates that vegetative to reproductive fungus Xylariales sp. PSU-ES163. Natural Product phase transition of meristem is regulated by sugar Research: Formerly Natural Product Letters, 30(1), balance. Functional Plant Biology, 42(6), 589-598. 46-51.
2. Akkasaeng, C., Tantisuwichwong, N., Ngamhui, N., 10. Aryuman, P., Lertsiri, S., Visessanguan, W., Niamsiri, Roytrakul, S., Jogloy, S. and Pathanothai, A. (2015). N., Bhumiratana, A. and Assavanig, A. (2015). Changes in Protein Expression in Peanut Leaves in the Glutaminase-producing Meyerozyma (Pichia) Response to Progressive Water Stresse. Pakistan guilliermondii isolated from Thai soy sauce Journal of Biological Sciences, 18(1), 19-26. fermentation. International Journal of Food Microbiology, 192, 7–12. 3. Aksoy, M., Pootakham, W. and Grossman, A.R. (2014). Critical Function of a Chlamydomonas reinhardtii 11. Asasutjarit, R., Theerachayanan, T., Kewsuwan, P., Putative Polyphosphate Polymerase Subunit during Veeranodha, S., Fuongfuchat, A. and Ritthidej, G.C. Nutrient Deprivation. Plant Cell, 26(8), 1-16. (2015). Development and Evaluation of Diclofenac Sodium Loaded-N-Trimethyl Chitosan Nanoparticles 4. Amornbunchornvej, C., Intarapanich, A., Assawamakin, for Ophthalmic Use. AAPS PharmSciTech, 16(5) 1013- A. and Tongsima, S. (2014). iNJclust: Iterative Neighbor- 1024. Joining Tree Clustering Framework for Inferring Population Structure. IEEE-ACM Transactions on 12. Beseli, A., Amnuaykanjanasin, A., Herrero, S., Thomas, Computational Biology and Bioinformatics, 11(5), E. and Daub, M.E. (2015). Membrane transporters in 903-914. self resistance of Cercospora nicotianae to the photoactivated toxin cercosporin. Current Genetics, 5. Ananphongmanee, V., Srisala, J., Sritunyalucksana, K. 61(4), 601-620. and Boonchird, C. (2015). Yeast Surface Display of two Proteins Previously Shown to Be Protective Against 13. Bhunchoth, A., Phironrit, N., Leksomboon, C., White Spot Syndrome Virus (WSSV) in Shrimp. PLOS Chatchawankanphanich, O., Kotera, S., Narulita, E., ONE, 10(6), e0128764. Kawasaki, T., Fujie, M. and Yamada, T. (2015). Isolation of Ralstonia solanacearum-infecting bacteriophages 6. Apitanyasai, K., Amparyup, P., Charoensapsri, W., from tomato fields in Chiang Mai, Thailand, and their Senapin, S. and Tassanakajon, A. (2015). Role of experimental use as biocontrol agents. Journal of Penaeus monodon hemocyte homeostasis associated Applied Microbiology, 118(4), 1023-1033. protein (PmHHAP) in regulation of caspase-mediated apoptosis. Developmental and Comparative 14. Boonkhot, P., Tadee, P., Yamsakul, P., Pocharoen, C., Immunology, 53(1), 234–243. Chokesajjawatee, N. and Patchanee, P. (2015). Class 1 integrons characterization and multilocus sequence 7. Ardhan, N., Ruttithiwapanich, T., Songkasiri, W. and typing of Salmonella spp. from swine production Phalakornkule, C. (2015). Comparison of performance chains in Chiang Mai and Lamphun provinces, of continuous-flow and batch electrocoagulators: A Thailand. Japanese Journal of Veterinary Research, case study for eliminating reactive blue 21 using iron 63(2), 83-94. electrodes. Separation and Purification Technology, 146, 75-84. 15. Buaklin, A., Jantee, N., Sittikankaew, K., Chumtong, P., Janpoom, S., Menasveta, P., Klinbunga, S. and 8. Arunpanichlert, J., Rukachaisirikul, V., Phongpaichit, Khamnamtong, B. (2015). Expression and S., Supaphon, O. and Sakayaroj, J. (2015). polymorphism of farnesoic acid O-methyltransferase Meroterpenoid, isocoumarin and phenol derivatives (FAMeT) and association between its SNPs and from the seagrass-derived fungus Pestalotiopsis sp. reproduction-related parameters of the giant tiger PSU-ES194. Tetrahedron, 71(5), 882-888. shrimp Penaeus monodon. Aquaculture, 441, 106-117. Appendices 65
16. Bunbamrung, N., Intaraudom, C., Supothina, S., 24. Charoenyingcharoen, P., Matsutani, M., Yakushi, T., Komwijit, S. and Pittayakhajonwut, P. (2015). Theeragool, G., Yukphan, P. and Matsushita, K. (2015). Antibacterial and anti-phytopathogenic substances A functionally critical single nucleotide polymorphism from the insect pathogenic fungus Gibellula sp. in the gene encoding the membrane-bound alcohol BCC36964. Phytochemistry Letters, 12, 142–147. dehydrogenase found in ethanol oxidation-deficient Gluconobacter thailandicus. Gene, 567(2), 201–207. 17. Bunterngsook, B., Eurwilaichitr, L., Thamchaipenet, A. and Champreda, V. (2015). Binding characteristics and 25. Chitnumsub, P., Jaruwat, A., Riangrungroj, P., Ittarat, synergistic effects of bacterial expansins on cellulosic W., Noytanom, K., Oonanant, W., Vanichthanankul, J., and hemicellulosic substrates. Bioresource Technology, Chuankhayan, P., Maenpuen, S., Chen, C.J., Chaiyen, 176, 129-135. P., Yuthavong, Y. and Leartsakulpanich, U. (2014). Structures of Plasmodium vivax serine 18. Bunyapaiboonsri, T., Yoiprommarat, S., Nopgason, R., hydroxymethyltransferase: implications for ligand- Intereya, K., Suvannakad, R. and Sakayaroj, J. (2015). binding specificity and functional control. Acta Palmarumycins from the mangrove fungus BCC 25093. Crystallographica Section D Biological Crystallography, Tetrahedron, 71(34), 5572–5578. 70(12), 3177-3186. 19. Chaikaew, S., Tepkasikul, P., Young, G.M., Osako, K., 26. Chruewkamlow, N., Pata, S., Mahasongkrama, K., Benjakul, S. and Visessanguan, W. (2015). Fixed-bed Laopajon, W., Kasinrerk, W. and Chiampanichayakul, degradation of histamine in fish sauce by immobilized S. (2015). β3 subunit of Na,K ATPase regulates T cell whole cells of Natrinema gari BCC 24369. Fisheries activation with no involvement of Na,K ATPase activity. Science, 81(5), 971-981. Immunobiology, 220(5), 634-640. 20. Chaikhumwang, P., Tantituvanont, A., Tripipat, T., 27. Daengrot, C., Rukachaisirikul, V., Tansakul, C., Tipsombatboon, P., Piriyapongsa, J. and Nilubol, D. Thongpanchang, T., Phongpaichit, S., Bowornwiriyapan, (2015). Dynamics and evolution of highly pathogenic K. and Sakayaroj, J. (2015). Eremophilane porcine reproductive and respiratory syndrome virus Sesquiterpenes and Diphenyl Thioethers from the Soil following its introduction into a herd concurrently Fungus Penicillium copticola PSU-RSPG138. Journal of F infected with both types 1 and 2. Infection Genetics Natural Products, 78(4), 615-622. and Evolution, 30, 164–174. 28. Dechtawewat, T., Songprakhon, P., Limjindaporn, T., 21. Chanama, M., Thongkrachang, N., Suriyachadkun, C. Puttikhunt, C., Kasinrerk, W., Saitornuang, S., and Chanama, S. (2015). Kutzneria chonburiensis sp. Yenchitsomanus, P-T. and Noisakran, S. (2015). Role nov., isolated from soil. International Journal of of human heterogeneous nuclear ribonucleoprotein Systematic and Evolutionary Microbiology, 65, 4169- C1/C2 in dengue virus replication. Virology Journal, 4174. 12, 14. 22. Charoenrat, T., Sangprapai, K., Promdonkoy, P., 29. Deelai, S., Suetrong, S., Damrianant, S., Unagul, P. and Kocharin, K., Tanapongpipat, S. and Roongsawang, N. Sakkayawong, N. (2015). Isolation and identification (2015). Enhancement of thermostable β-glucosidase of native lower fungi for polyunsaturated fatty acid production in a slow methanol utilization strain of (PUFA) production in Thailand, and the effect of carbon Pichia pastoris by optimization of the specific methanol and nitrogen sources on growth and production. supply rate. Biotechnology and Bioprocess Engineering, African Journal of Biotechnology, 14(17), 1449-1460. 20(2), 315-323. 30. Dejnirattisai, W., Wongwiwat, W., Supasa, S., Zhang, 23. Charoensapsri, W., Sangsuriya, P., Lertwimol, T., X., Dai, X., Rouvinsky, A., Jumnainsong, A., Edwards, Gangnonngiw, W., Phiwsaiya, K. and Senapin, S. (2015). C., Quyen, N.T.H., Duangchinda, T., Grimes, J.M., Tsai, Laminin receptor protein is implicated in hemocyte W.Y., Lai, C.H., Wang, W.K., Malasit, P., Farrar, J., homeostasis for the whiteleg shrimp Penaeus Simmons, C.P., Zhou, Z.H., Rey, F.A., Mongkolsapaya, (Litopenaeus) vannamei. Developmental and J. and Screaton, G.R. (2015). A new class of highly Comparative Immunology, 51(1), 39-47. potent, broadly neutralizing antibodies isolated from viremic patients infected with dengue virus. Nature Immunology, 16, 170-177. 66 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
31. Disratthakit, A., Meada, S., Prammananan, T., 39. Havanapan, P., Taengchaiyaphum, S., Ketterman, A.J. Thaipisuttikul, I., Doi, N. and Chaiprasert, A. (2015). and Krittanai, C. (2016). Yellow head virus infection in Genotypic diversity of multidrug-, quinolone- and black tiger shrimp reveals specific interaction with extensively drug-resistant Mycobacterium tuberculosis granule-containing hemocytes and crustinPm1 as a isolates in Thailand. Infection Genetics and Evolution, responsive protein. Developmental and Comparative 32, 432-439. Immunology, 54(1), 126–136.
32. Dokladda, K., Billamas, P. and Palittapongarnpim, P. 40. Hotaka, D., Amnuaykanjanasin, A., Maketon, C., (2015). Different behaviours of promoters in Siritutsoontorn, S. and Maketon, M. (2015). Efficacy Mycobacterium tuberculosis H37Rv and H37Ra. World of Purpureocillium lilacinum CKPL-053 in controlling Journal of Microbiology and Biotechnology, 31(2), Thrips palmi (Thysanoptera: Thripidae) in orchid farms 407-413. in Thailand. Applied Entomology and Zoology, 50(3), 317–329. 33. Dong, H. T., Senapin, S., Frentz, B. L. and Rodkhum, C. (2015). Virulence assay of rhizoid and non-rhizoid 41. Hua, Y., Ekkhara, W., Sansenya, S., Srisomsap, C., morphotypes of Flavobacterium columnare in red Roytrakul, S., Saburi, W., Takeda, R., Matsuura, H., tilapia,Oreochromis sp., fry. Journal of Fish Diseases, Mori, H. and Cairns, J.R.K. (2015). Identification of rice doi: 10.1111/jfd.12385. Os4BGlu13 as a β-glucosidase which hydrolyzes gibberellin A4 1-O-β-D-glucosyl ester, in addition to 34. Dong, H.T., Nguyen, V.V., Le, H.D., Sangsuriya, P., tuberonic acid glucoside and salicylic acid derivative Jitrakorn, S., Saksmerprome, V., Senapin, S. and glucosides. Archives of Biochemistry and Biophysics, Rodkhum, C. (2015). Naturally concurrent infections 583, 36-46. of bacterial and viral pathogens in disease outbreaks in cultured Nile tilapia (Oreochromis niloticus) farms. 42. Hunsawong, T., Sunintaboon, P., Warit, S., Aquaculture, 448, 427–435. Thaisomboonsuk, B., Jarman, R.G., Yoon, I-K., Ubol, S. and Fernandez, S. (2015). A novel dengue virus 35. Dong, H-T., Nguyen, V-V., Phiwsaiya, K., Gangnonngiw, serotype-2 nanovaccine induces robust humoral and W., Withyachumnarnkul, B., Rodkhum, C. and Senapin, cell-mediated immunity in mice. Vaccine, 33(14), S. (2015). Concurrent infections of Flavobacterium 1702-1710. columnare and Edwardsiella ictaluri in striped catfish, Pangasianodon hypophthalmus in Thailand. 43. Hunsawong, T., Sunintaboon, P., Warit, S., Aquaculture, 448, 142-150. Thaisomboonsuk, B., Jarman, R.G., Yoon, I-K., Ubol, S. and Fernandez, S. (2015). Immunogenic Properties of 36. Gadaj, A., Cooper, K.M., Karoonuthaisiri, N., Furey, A. a BCG Adjuvanted Chitosan Nanoparticle-Based and Danaher, M. (2015). Determination of the Dengue Vaccine in Human Dendritic Cells. PLOS persistence of dimetridazole, metronidazole and Neglected Tropical Disease, 9(9), e0003958. ronidazole residues in black tiger shrimp (Penaeus monodon) tissue and their stability during cooking. 44. Imman, S., Arnthong, J., Burapatana, V., Champreda, Food additives & contaminants. Part A, Chemistry, V. and Laosiripojana, N. (2015). Fractionation of rice analysis, control, exposure & risk assessment, 32(2), straw by a single-step solvothermal process: Effects 180-193. of solvents, acid promoters, and microwave treatment. Renewable Energy, 83, 663–673. 37. Grajeda-Iglesias, C., Salas, E., Barouh, N., Baréa, B., Panya, A., Figueroa-Espinoza, M.C. (2015). Antioxidant 45. Imman, S., Arnthong, J., Burapatana, V., Champreda, activity of protocatechuates evaluated by DPPH, ORAC, V. and Laosiripojana, N. (2015). Influence of alkaline and CAT methods. Food Chemistry, 194, 749-757. catalyst addition on compressed liquid hot water pretreatment of rice straw. Chemical Engineering 38. Haque, M.F., Boonhok, R., Prammananan, T., Journal, 278, 85-91. Chaiprasert, A., Utaisincharoen, P., Sattabongkot, J., Palittapongarnpim, P. and Ponpuak, M. (2015). 46. Intarasirisawat, R., Benjakul, S., Vissessanguan, W., Resistance to cellular autophagy by Mycobacterium Maqsood, S. and Osako, K. (2015). Skipjack roe protein tuberculosis Beijing strains. Innate Immunity, 21(7), hydrolysate combined with tannic acid increases the 746-758. stability of fish oil upon microencapsulation. European Journal of Lipid Science and Technology, 117(5), 646–656. Appendices 67
47. Intasai, N., Pata, S., Tragoolpua, K. and Tayapiwatana, 55. Janvilisri, T., Leelawat, K., Roytrakul, S., Paemanee, A. C. (2015). Recombinant Multivalent EMMPRIN and Tohtong, R. (2015). Novel Serum Biomarkers to Extracellular Domain Induces U937 Human Leukemia Differentiate Cholangiocarcinoma from Benign Biliary Cell Apoptosis by Downregulation of Monocarboxylate Tract Diseases Using a Proteomic Approach. Disease Transporter 1 and Activation of Procaspase-9.Applied Markers, 2015, Article ID 105358. Biochemistry and Biotechnology, 176(6), 1781-1790. 56. Jearaphunt, M., Amparyup, P., Sangsuriya, P., 48. Isaka, M., Chinthanom, P., Rachtawee, P., Srichomthong, Charoensapsri, W., Senapin, S. and Tassanakajon, A. K., Srikitikulchai, P., Kongsaeree, P. and Prabpai, S. (2015). Shrimp Serine Proteinase Homologues (2015). Cytotoxic hydroanthraquinones from the PmMasSPH-1 and -2 Play a Role in the Activation of mangrove-derived fungus Paradictyoarthrinium the Prophenoloxidase System. PLOS ONE, 10(3), diffractum BCC 8704. Journal of Antibiotics, 68(5), e0121073. 334-338. 57. Jeennor, S., Cheawchanlertfa, P., Suttiwattanakul, S., 49. Isaka, M., Haritakun, R., Supothina, S., Choowong, W. Panchanawaporn, S., Chutrakul, C. and Laoteng, K. and Mongkolsamrit, S. (2014). N-Hydroxypyridone (2015). The codon-optimized Δ 6 -desaturase gene of alkaloids, chromone derivatives, and Pythium sp. as an empowering tool for engineering tetrahydroxanthones from the scale-insect pathogenic n3/n6 polyunsaturated fatty acid biosynthesis. BMC fungus Orbiocrella sp. BCC 33248. Tetrahedron, 70(48), Biotechnology, 15, 82. 9198–9203. 58. Jengarn, J., Wongthida, P., Wanasen, N., Frantz, P.N., 50. Isaka, M., Palasarn, S., Sappan, M., Srichomthong, K., Wanitchang, A. and Jongkaewwattana, A. (2015). Karunarathna, S.C. and Hyde, K.D. (2015). Genetic manipulation of porcine epidemic diarrhea Prenylhydroquinone-Derived Secondary Metabolites virus recovered from a full-length infectious cDNA from Cultures of the Basidiomycete Lentinus similis clone. Journal of General Virology, 96(8), 2206-2218. BCC 52578. Natural Product Communications, 10(8), 59. Jindamorakot, S., Am-In, S., Kaewwichian, R. and 1391-1393. Limtong, S. (2015). Yamadazyma insecticola f.a., sp. F51. Isaka, M., Yangchum, A., Supothina, S., Boonpratuang, nov. and Yamadazyma epiphylla f.a., sp. nov., two novel T., Choeyklin, R., Kongsaeree, P. and Prabpai, S. (2015). yeast species. International Journal of Systematic and Aromadendrane and cyclofarnesane sesquiterpenoids Evolutionary Microbiology, 65(Pt 4), 1290-1296. from cultures of the basidiomycete Inonotus sp. BCC 60. Jirakkakul, J., Cheevadhanarak, S., Punya, J., Chutrakul, 23706. Phytochemistry, 118, 94-101. C., Senachak, J., Buajarern, T., Tanticharoen, M. and 52. Isaka, M., Yangchum, A., Supothina, S., Laksanacharoen, Amnuaykanjanasin, A. (2015). Tenellin acts as an iron P., Luangsa-ard, J.J. and Hywel-Jones, N.L. (2015). chelator to prevent iron-generated reactive oxygen Ascochlorin derivatives from the leafhopper pathogenic species toxicity in the entomopathogenic fungus fungus Microcera sp. BCC 17074. Journal of Antibiotics, Beauveria bassiana. FEMS Microbiology Letters, 68, 47–51. 362(2), 1-8.
53. Jaidee, A., Promchai, T., Trisuwan, K., Laphookhieo, S., 61. Jones, E.B.G., Suetrong, S., Sakayaroj, J., Bahkali, A.H., Rattanajak, R., Kamchonwongpaisan, S., Pyne, S.G. Abdel-Wahab, M.A., Boekhout, T. and Pang, K.L. and Ritthiwigrom, T. (2015). Cytotoxic and antimalarial (2015). Classification of marine Ascomycota, alkaloids from the twigs of Dasymaschalon Basidiomycota, Blastocladiomycota and obtusipetalum. Natural Product Communications, Chytridiomycota. Fungal Diversity, 73(1), 1-72. 10(7), 1175-1178. 62. Juntawong, P., Sirikhachornkit, A., Pimjan, R., 54. Jannoey, P., Pongprasert, W., Lumyong, S., Roytrakul, Sonthirod, C., Sangsrakru, D., Yoocha, T., S. and Nomura, M. (2015). Comparative proteomic Tangphatsornruang, S. and Srinives, P. (2014). analysis of two rice cultivars (Oryza sativa L.) contrasting Elucidation of the molecular responses to waterlogging in Brown Planthopper (BPH) stress resistance. Plant in Jatropha roots by transcriptome profiling.Frontiers Omics, 8(2), 96-105. in Plant Science, 5, 658. 68 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
63. Kaewseekhao, B., Naranbhai, V., Roytrakul, S., Namwat, 71. Khunnamwong, P., Surussawadee, J., Jindamorakot, W., Paemanee, A., Lulitanond, V., Chaiprasert, A. and S., Ribeiro, J.R.A., Hagler, A.N. and Limtong, S. (2015). Faksri, K. (2015). Comparative Proteomics of Activated Occultifur tropicalis f.a., sp. nov., a novel THP-1 Cells Infected with Mycobacterium tuberculosis cystobasidiomycetous yeast species isolated from Identifies Putative Clearance Biomarkers for tropical regions. International Journal of Systematic Tuberculosis Treatment. PLOS ONE, 10(7), e0134168. and Evolutionary Microbiology, 65, 1578-1582.
64. Kaewwichian, R., Jindamorakot, S., Am-In, S., Sipiczki, 72. Khunrattanaporn, N., Rijiravanich, P., Somasundrum, M. and Limtong, S. (2015). Hannaella siamensis sp. M. and Surareungchai, W. (2015). Highly sensitive nov. and Hannaella phetchabunensis sp. nov., two new electrochemical detection of genomic DNA based on anamorphic basidiomycetous yeast species isolated stem loop probes structured for magnetic collection from plants. International Journal of Systematic and and measurement via metalised hollow polyelectrolyte Evolutionary Microbiology, 65(Pt 4), 1297-1303. shells. Biosensors and Bioelectronics, 73, 181-187.
65. Kamolsukyumyong, W., Vanavichit, A., Sookwong, P. 73. Kittipongpittaya, K., Panya, A., Cui, L., McClements, and Mahatheeranont, S. (2014). Monoterpenoid D.J. and Decker, E.A. (2014). Association Colloids Allelochemicals in Resistance Rice Varieties against Formed by Multiple Surface Active Minor Components Brown Planthoppers, Nilaparvata Lugens (Stål). and Their Effect on Lipid Oxidation in Bulk Oil.Journal Journal of Advanced Agricultural Technologies, 1(2), of the American Oil Chemists Society, 91(11), 1955- 82-88. 1965.
66. Kamphee, H., Chaiprasert, A., Prammananan, T., 74. Klaysuban, A., Sakayaroj, J. and Jones, E.B.G. (2014). Wiriyachaiporn, N., Kanchanatavee, A. and Dharakul, An additional marine fungal lineage in the Diatrypaceae, T. (2015). Rapid Molecular Detection of Multidrug- Xylariales: Pedumispora rhizophorae. Botanica Resistant Tuberculosis by PCR-Nucleic Acid Lateral Marina, 57(5), 413-420. Flow Immunoassay. PLOS ONE, 10(9), e0137791. 75. Kobmoo, N., Mongkolsamrit, S., Wutikhun, T., 67. Karoonuthaisiri, N., Charlermroj, R., Teerapornpuntakit, Tasanathai, K., Khonsanit, A., Thanakitpipattana, D. J., Kumpoosiri, M., Himananto, O., Grant, I.R., and Luangsa-Ard, J.J. (2015). New Species of Gajanandana, O. and Elliott, C.T. (2015). Bead array Ophiocordyceps unilateralis, an ubiquitous pathogen for Listeria monocytogenes detection using specific of ants from Thailand. Fungal Biology, 119(1), 44-52. monoclonal antibodies. Food Control, 47, 462–471. 76. Kongprapan, T., Rukachaisirikul, V., Saithong, S., 68. Khamlor, T., Pongpiachan, P., Parnpai, R., Punyawai, Phongpaichit, S., Poonsuwan, W. and Sakayaroj, J. K., Sangsritavong, S. and Chokesajjawatee, N. (2015). (2015). Cytotoxic cytochalasins from the endophytic Bovine embryo sex determination by multiplex loop- fungus Eutypella scoparia PSU-H267. Phytochemistry mediated isothermal amplification. Theriogenology, Letters, 13, 171–176. 83(5), 891-896. 77. Kongyen, W., Rukachaisirikul, V., Phongpaichit, S. and 69. Khaokhajorn, P., Samipak, S., Nithithanasilp, Sakayaroj, J. (2015). A new hydronaphthalenone from S.,Tanticharoen, M. and Amnuaykanjanasin, A. (2015). the mangrove-derived Daldinia eschscholtzii PSU- Production and secretion of naphthoquinones is STD57. Natural Product Research. Formerly Natural mediated by the MFS transporter MFS1 in the Product Letters, 29(21), 1995-1999. entomopathogenic fungus Ophiocordyceps sp. 78. Kornsakulkarn, J., Saepua, S., Srijomthong, K., BCC1869. World Journal of Microbiology and Rachtawee, P. and Thongpanchang, C. (2015). Biotechnology, 31(10), 1543-1554. Quinazolinone alkaloids from actinomycete 70. Khemkhao, M., Techkarnjanaruk, S. and Phalakornkule, Streptomyces sp. BCC 21795. Phytochemistry Letters, C. (2015). Simultaneous treatment of raw palm oil mill 12, 6-8. effluent and biodegradation of palm fiber in a high-rate CSTR. Bioresource Technology, 177, 17-27. Appendices 69
79. Kotawong, K., Thitapakorn, V., Roytrakul. S., 87. Liu, J.K., Hyde, K.D. Jones, E. B. G., Ariyawansa, H.A., Phaonakrop, N., Viyanant, V. and Na-Bangchang, K. Bhat, D.J., Boonmee, S., Maharachchikumbura, S.S.N., (2015). Plasma Phosphoproteome and Differential McKenzie, E.H.C., Phookamsak, R., Phukhamsakda, C., Plasma Phosphoproteins with Opisthorchis Viverrini- Shenoy, B.D., Abdel-Wahab, M.A., Buyck, B., Chen, J., Related Cholangiocarcinoma. Asian Pacific Journal of Chethana, K.W.T., Singtripop, C., Dai, D.Q., Dai, Y.C., Cancer Prevention, 16(3), 1011-1018. Daranagama, D.A., Dissanayake, A.J. Doilom, M., D’souza, M.J., Fan, X.L., Goonasekara, I.D., Hirayama, 80. Kriangphan, N., Vuttipongchaikij, S., Kittiwongwattana, K., Hongsanan, S., Jayasiri, S.C., Jayawardena, R.S., C., Suttangkakul, A., Pinmanee, P., Sakulsathaporn, A., Karunarathna, S.C., Li, W.J., Mapook, A., Norphanphoun, Suwimon, R., Suputtitada, S., Chanvivattana, Y. and C., Pang, K.L., Perera, R.H., Peršoh, D., Pinruan, U., Apisitwanich, S. (2015). Effects of Sequence and Senanayake, I.C., Somrithipol, S., Suetrong, S., Tanaka, Expression of Eight Anthocyanin Biosynthesis Genes K., Thambugala, K.M., Tian, Q., Tibpromma, S., on Floral Coloration in Four Dendrobium Hybrids. Udayanga, D., Wijayawardene, N.N., Wanasinghe, D., Journal of the Japanese Society for Horticultural Wisitrassameewong, K., Zeng, X.Y., Abdel-Aziz, F.A., Science, 84(1), 83-92. Adamčík, S., Bahkali, A.H., Boonyuen, N., Bulgakov, T., 81. Krongsin, J., Gamonpilas, C., Methacanon, P., Panya, Callac, P., Chomnunti, P., Greiner, K., Hashimoto, A., A. and Goh, S.M. (2015). On the stabilisation of Hofstetter, V., Kang, J.C., Lewis, D., Li, X.H., Liu, X.Z., calcium-fortified acidified soy milks by pomelo pectin. Liu, Z.Y., Matsumura, M., Mortimer, P.E., Rambold, G., Food Hydrocolloids, 50, 128-136. Randrianjohany, E., Sato, G., Sri-Indrasutdhi, V., Tian, C.M., Verbeken, A., Brackel, W.V., Wang, Y., Wen, T.C., 82. Kueanjinda, P., Roytrakul, S. and Palaga, T. (2015). A Xu, J.C., Yan, J.Y., Zhao, R.L., Camporesi, E. (2015). Novel Role of Numb as A Regulator of Pro-inflammatory Fungal diversity notes 1–110: taxonomic and Cytokine Production in Macrophages in Response to phylogenetic contributions to fungal species. Fungal Toll-like Receptor 4. Scientific Reports, 5, 12784. Diversity, 72(1), 1-197.
83. Kurdi, P. and Hansawasdi, C. (2015). Assessment of 88. Lomthong, T., Chotineeranat, S. and Kitpreechavanich, the prebiotic potential of oligosaccharide mixtures V. (2015). Production and characterization of raw F from rice bran and cassava pulp. Lwt-Food Science and starch degrading enzyme from a newly isolated Technology, 63(2), 1288–1293. thermophilic filamentous bacterium, Laceyella sacchari LP175. Starch/Stärke, 67(3-4), 255-266. 84. Laothanachareon, T., Bunterngsook, B., Suwannarangsee, S., Eurwilaichitr, L. and Champreda, 89. Low, K.F., Rijiravanich, R., Singh, K.K.B., Surareungchai, V. (2015). Synergistic action of recombinant accessory W. and Yean, C.Y. (2015). An Electrochemical hemicellulolytic and pectinolytic enzymes to Genosensing Assay Based on Magnetic Beads and Gold Trichoderma reesei cellulase on rice straw degradation. Nanoparticle-Loaded Latex Microspheres for Vibrio Bioresource Technology, 198, 682–690. cholerae Detection. Journal of Biomedical Nanotechnology, 11(4), 702-710(9). 85. Lekakarn, H., Promdonkoy, B. and Boonserm, P. (2015). Interaction of Lysinibacillus sphaericus binary toxin 90. Maenpuen, S., Amornwatcharapong, W., Krasatong, with mosquito larval gut cells: Binding and P., Sucharitakul, J., Palfey, B.A., Yuthavong, Y., internalization.Journal of Invertebrate Pathology, 132, Chitnumsub, P., Leartsakulpanich, U. and Chaiyen, P. 125–131. (2015). Kinetic Mechanism and the Rate-Limiting Step of Plasmodium vivax Serine Hydroxymethyltransferase. 86. Lerksuthirat, T., Lohnoo, T., Inkomlue, R., Rujirawat, T., Journal of Biological Chemistry, 290(13), 8656-8665. Yingyong, W., Khositnithikul, R., Phaonakrop, N., Roytrakul, S., Sullivan, T.D. and Krajaejun, T. (2015). The Elicitin-Like Glycoprotein, ELI025, Is Secreted by the Pathogenic Oomycete Pythium insidiosum and Evades Host Antibody Responses. PLOS ONE, 10(3), e0118547. 70 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
91. Maharachchikumbura, S.S.N., Hyde, K.D.,Gareth Jones, 98. Mitupatum, T., Aree, K., Kittisenachai, S., Roytrakul, E. B., McKenzie, E.H.C., Huang, S.K., Abdel-Wahab, M. S., Puthong, S., Kangsadalampai, S. and Rojpibulstit, A., Daranagama, D. A., Dayarathne, M., D’souza, M. J., P. (2015). Hep88 mAb-Mediated Paraptosis-Like Goonasekara, I.D., Hongsanan, S., Jayawardena, R.S., Apoptosis in HepG2 Cells via Downstream Upregulation Kirk, P.M., Konta, S., Liu, J.K., Liu, Z.Y., Norphanphoun, and Activation of Caspase-3, Caspase-8 and Caspase-9. C., Pang, K.L., Perera, R.H., Senanayake, I.C., Shang, Asian Pacific Journal of Cancer Prevention, 16(5), 1771- Q., Shenoy, B.D., Xiao, Y., Bahkali, A.H., Kang, J., 1779. Somrothipol, S., Suetrong, S., Wen, T. and Xu, J. (2015). 99. Miyashita, M., Yukphan, P., Chaipitakchonlatarn, W., Towards a natural classification and backbone tree for Malimas, T., Sugimoto, M., Yoshino, M., Kamakura, Y., Sordariomycetes. Fungal Diversity, 72(1), 199-301. Potacharoen, W., Tanasupawat, S., Tanaka, N., 92. Mahasongkram, K., Pata, S., Chruewkamlow, N. and Nakagawa, Y. and Suzuki, K-I. (2015). Lactobacillus Kasinrerk, W. (2015). Identification of a T cell surface plajomi sp. nov. and Lactobacillus modestisalitolerans molecule using a monoclonal antibody produced by sp. nov., isolated from traditional fermented foods. TCR/CD3 complex immunization.Asian Pacific Journal International Journal of Systematic and Evolutionary of Allergy and Immunology, 33(2). Microbiology, 65(8), 2485-2490.
93. Malila, Y., Srimarut, Y., U-chupaj, J., Strasburg, G. and 100. Mokmak, W., Chunsrivirot, S., Hannongbua, S., Visessanguan, W. (2015). Monitoring of Chicken RNA Yuthavong, Y., Tongsima, S. and Kamchonwongpaisan, Integrity as A Function of Prolonged Postmortem S. (2014). Molecular Dynamics of Interactions between Duration. Asian-Australasian Journal of Animal Rigid and Flexible Antifolates and Dihydrofolate Sciences, 28(11),1649-1656. Reductase from Pyrimethamine-Sensitive and Pyrimethamine–Resistant Plasmodium falciparum. 94. Mamimin, C., Singkhala, A., Kongjan, P., Suraraksa, B., Chemical Biology and Drug Design, 84(4), 450–461. Prasertsan, P., Imai, T. and O-Thong, S. (2015). Two- stage thermophilic fermentation and mesophilic 101. Mongkolsamrit, S., Khonsanit, A., Noisripoom, W., methanogen process for biohythane production from Baldorj, P. and Luangsa-Ard, J. J. (2014). Aschersonia palm oil mill effluent.International Journal of Hydrogen narathiwatensis sp. nov. from southern Thailand. Energy, 40(19), 6319–6328. Mycotaxon, 129(1), 33-40.
95. McConkey, K.M., Brockelman, W.Y. and Saralamba, C. 102. Moonrin, N., Songtawee, N., Rattanabunyong, S., (2014). Mammalian Frugivores With Different Foraging Chunsrivirot, S., Mokmak, W., Tongsima, S. and Behavior Can Show Similar Seed Dispersal Effectiveness. Choowongkomon, K. (2015). Understanding the Biotropica, 46(6), 647–651. molecular basis of EGFR kinase domain/MIG-6 peptide recognition complex using computational analyses. 96. Mhuantong, W., Charoensawan, V., Kanokratana, P., BMC Bioinformatics, 16, 103. Tangphatsornruang, S. and Champreda, V. (2015). Comparative analysis of sugarcane bagasse 103. Mukkata, K., Kantachote, D., Wittayaweerasak, B., metagenome reveals unique and conserved biomass- Techkarnjanaruk, S. and Boonapatcharoen, N. (2015). degrading enzymes among lignocellulolytic microbial Diversity of purple nonsulfur bacteria in shrimp ponds communities. Biotechnology for Biofuels, 8, 16. with varying mercury levels. Saudi Journal of Biological Sciences, doi:10.1016/j.sjbs.2015.05.014. 97. Mhuantong, W., Wongwilaiwalin, S., Laothanachareon, T., Eurwilaichitr, L, Tangphatsornruang, S., 104. Mukkata, K., Kantachote, D., Wittayaweerasak, B., Boonchayaanant, B., Limpiyakorn, T., Pattaragulwanit, Techkarnjanaruk, S., Mallavarapu, M. and Naidu, R. K., Punmatharith, T., McEvoy, J., Khan, E., Rachakornkij, (2015). Distribution of Mercury in Shrimp Ponds and M. and Champreda, V. (2015). Survey of Microbial Volatilization of Hg by Isolated Resistant Purple Diversity in Flood Areas during Thailand 2011 Flood Nonsulfur Bacteria. Water Air and Soil Pollution, 226, Crisis Using High-Throughput Tagged Amplicon 148. Pyrosequencing. PLOS ONE, 10(5), e0128043. Appendices 71
105. Namvongsakool, P., Asuvapongpatana, S., Senapin, S., 114. Obchoei, S., Sawanyawisuth, K., Wongkham, C., Weerachatyanukul, W., Treerattrakool, S. and Kasinrerk, W., Yao, Q., Chen, C. and Wongkham, S. Withyachumnarnkul, B. (2015). A novel localization of (2015). Secreted cyclophilin A mediates G1/S phase molt-inhibiting hormone in the tegumental glands of transition of cholangiocarcinoma cells via CD147/ shrimp Penaeus monodon and its possible role in ERK1/2 pathway. Tumor Biology, 36(2), 849-859. shrimp molting. Aquaculture, 438, 129-137. 115. OO, K-S., Kongjaimun, A., Khanthong, S., Yi, M., Myint, 106. Narkpuk, J., Jaru-Ampornpan, P., Subali, T., Bertulfo, T-T., Korinsak, S., Siangliw, J-L., Myint, K-M., Vanavichit, F.C.T., Wongthida, P. and Jongkaewwattan, A. (2015). A., Malumpong, C. and Toojinda, T. (2015). Mechanistic study of intertypic nucleoprotein complex Characterization of Myanmar Paw San Hmwe formation and its inhibitory effect toward influenza A Accessions Using Functional Genetic Markers. Rice virus. Virology, 485, 104–115. Science, 22(2), 3.
107. Nawattanapiaboon, K., Kiatpathomchai, W., 116. Panichakul, T., Ponnikorn, S., Roytrakul, S., Paemanee, Santanirand, P., Vongsakulyanon, A., Amarit, R., A., Kittisenachai, S., Hongeng, S. and Udomsangpetch, Somboonkaew, A., Sutapun, B. and Srikhirin, T. (2015). R. (2015). Plasmodium vivax inhibits erythroid cell SPR-DNA array for detection of methicillin-resistant growth through altered phosphorylation of the Staphylococcus aureus (MRSA) in combination with cytoskeletal protein ezrin. Malaria Journal, 14, 138. loop-mediated isothermal amplification. Biosensors 117. Panijpan, B., Kowasupat, C., Laosinchai, P., Ruenwongsa, and Bioelectronics, 74, 335–340. P., Phongdara, A., Senapin, S., Wanna, W., Phiwsaiya, 108. Naziri, D., Quaye, W., Siwoku, B., Wanlapatit, S., Phu, K., Kühne, J. and Fasquel, F. (2014). Southeast Asian T.V. and Bennett, B. (2014). The diversity of postharvest mouth-brooding Betta fighting fish (Teleostei: losses in cassava value chains in selected developing Perciformes) species and their phylogenetic countries. Journal of Agriculture and Rural Development relationships based on mitochondrial COI and nuclear in the Tropics and Subtropics, 115(2), 111-123. ITS1 DNA sequences and analyses. Meta Gene, 2, 862–879. 109. Ngamhui, N., Tantisuwichwong, N., Roytrakul, S., Zhu, F Y. J., Li, Q. X. and Akkasaeng, C. (2015). Relationship 118. Panijpan, B., Laosinchai, P., Senapin, S., Kowasupat, between drought tolerance with activities of C., Ruenwongsa, P., Kühne, J. and Phiwsaiya, K. (2015). antioxidant enzymes in sugarcane. Indian Journal of Mitochondrial COI and nuclear RAG1 DNA sequences Plant Physiology, 20(2), 145–150. and analyses of specimens of the three morphologically established species in the genus Trichopsis 110. Ngamphiw, C., Tongsima, S. and Mutirangura, A. (Perciformes: Osphronemidae) reveal new/cryptic (2014). Roles of intragenic and intergenic L1s in mouse species. Meta Gene, 4, 17–28. and human. PLOS ONE, 9(11), e113434. 119. Panya, A., Temthawee, W., Phonsatta, N., Charoensuk, 111. Nguyen, V.V., Dong, H.T., Senapin, S., Pirarat. N. and D., Deetae, P., Visessanguan, W. and Decker, E.A. Rodkhum, C. (2015). Francisella noatunensis subsp. (2015). Apolar Radical Initiated Conjugated orientalis, an emerging bacterial pathogen affecting Autoxidizable Triene (ApoCAT) Assay: Effects of Oxidant cultured red tilapia (Oreochromis sp.) in Thailand. Locations on Antioxidant Capacities and Interactions. Aquaculture Research, doi:10.1111/are.12802. Journal of Agricultural and Food Chemistry, 63(34), 112. Nuchdang, S., Khemkhao, M., Techkarnjanaruk, S. and 7546–7555. Phalakornkule, C. (2015). Comparative biochemical 120. Panyakampol, J., Cheevadhanarak, S., Sutheeworapong, methane potential of paragrass using an unacclimated S., Chaijaruwanich, J., Senachak, J., Siangdung, W., and an acclimated microbial consortium.Bioresource Jeamton, W., Tanticharoen, M. and Paithoonrangsarid, Technology, 183, 111-119. K. (2015). Physiological and transcriptional responses 113. Nugraha, A.S., Hilou, A., Vandegraaff, N., Rhodes, D.I., to high temperature in Arthrospira (Spirulina) platensis Haritakun, R. and Keller, P.A. (2015). Bioactive C1. Plant and Cell Physiology, 56(3), 481-496. glycosides from the African medicinal plant Boerhavia erecta L. Natural Product Research. Formerly Natural Product Letters, 29(20), 1954-1958. 72 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
121. Paopang, P., Kasinrerk, W., Tayapiwatana, C., 128. Pitiwittayakul, N., Yukphan, P., Chaipitakchonlatarn, Seesuriyachan, P. and Butr-Indr, B. (2015). Multi- W., Yamada, Y. and Theeragool, G. (2015). Acetobacter Parameter Optimization Method and Enhanced thailandicus sp. nov., for a strain isolated in Thailand. Production of Secreted Recombinant Single Chain Annals of Microbiology, 65, 1855-1863. Variable Fragment against the HIV-1 P17 Protein From 129. Pitiwittayakul, N., Yukphan, P., Sintuprapa, W., Yamada, Escherichia Coli by Fed-Batch Fermentation. Y. and Theeragool, G. (2014). Identification of acetic Preparative Biochemistry and Biotechnology, doi:10. acid bacteria isolated in Thailand and assigned to the 1080/10826068.2015.1031388. genus Acetobacter by groEL gene sequence analysis. 122. Pattarapimol, T., Thuzar, M., Vanavichit, A., Annals of Microbiology, 65(3), 1557-1564. Tragoonrung, S., Roytrakul, S. and Jantasuriyarat, C. 130. Pootakham, W., Jomchai, N., Ruang-areerate, P., (2015). Identification of genes involved in somatic Shearman, J.R., Sonthirod, C., Sangsrakru, D., embryogenesis development in oil palm (Elaeis Tragoonrung, S. and Tangphatsornruang, S. (2015). guineensis Jacq.) using cDNA AFLP. Journal of Oil Palm Genome-wide SNP discovery and identification of QTL Research, 27(1), 1-11. associated with agronomic traits in oil palm using 123. Phanaksri, T., Luxananil, P., Panyim, S. and Tirasophon, genotyping-by-sequencing (GBS). Genomics, 105(5-6), W. (2015). Synergism of regulatory elements in σB- and 288-295. σA-dependent promoters enhances recombinant 131. Pootakham, W., Ruang-areerate, P., Jomchai, N., protein expression in Bacillus subtilis. Journal of Sonthirod, C., Sangsrakru, D., Yoocha, T., Bioscience and Bioengineering, 120(4), 470-475. Theerawattanasuk, K., Nirapathpongporn, K., 124. Phattanawiboon, B., Jariyapan, N., Roytrakul, S., Romruensukharom, P., Tragoonrung, S. and Paemanee, A., Sor-suwan, S., Intakhan, N., Chanmol, Tangphatsornruang, S. (2015). Construction of a high- W., Siriyasatien, P., Saeung, A. and Choochote, W. density integrated genetic linkage map of rubber tree (2014). Morphological and protein analyses of adult (Hevea brasiliensis) using genotyping-by-sequencing female salivary glands of Anopheles barbirostris (GBS). Frontiers in Plant Science, 6, 367. species A1 (Diptera: Culicidae). Tropical Biomedicine, 132. Pootakham, W., Shearman, J.R., Ruang-areerate, P., 31(4), 813-827. Sonthirod, C., Sangsrakru, D., Jomchai, N., Yoocha, T., 125. Pichyangkul, S., Yongvanitchit, K., Limsalakpetch, A., Triwitayakorn, K., Tragoonrung, S. and Kum-Arb, U., Im-Erbsin, R., Boonnak, K., Thitithayanont, Tangphatsornruang, S. (2014). Large-Scale SNP A., Jongkaewwattana, A., Wiboon-ut, S., Discovery through RNA Sequencing and SNP Mongkolsirichaikul, D., Mahanonda, R., Spring, M., Genotyping by Targeted Enrichment Sequencing in Chuang, I., Mason, C.J. and Saunders, D.L. (2015). Cassava (Manihot esculenta Crantz). PLOS ONE, 9(12), Tissue Distribution of Memory T and B Cells in Rhesus e116028. Monkeys following Influenza A Infection. Journal of 133. Prasertlux, S., Yocawibun, P., Janpoom, S., Klinbunga, Immunology, 195, 4378–4386. S., Menasveta, P. and Khamnamtong, B. (2015). 126. Piriyapongsa, J., Bootcha,i C., Ngamphiw, C. and Differential expression of X-box binding protein 1 Tongsima, S. (2014). microPIR2: a comprehensive during ovarian development and association between database for human-mouse comparative study of its SNP and growth-related parameters of the giant microRNA-promoter interactions.Database (Oxford), tiger shrimp Penaeus monodon. Aquaculture, 448, 2014, article ID bau115. 531–538.
127. Pitaksutheepong, C., Abhisingha, M., Dumnin, J. and 134. Prasertpornsakun, N., Raita, M., Laosiripojana, N. and Visessanguan, W. (2015). Isolation, detection and Champreda, V. (2015). Biocatalytic synthesis of starch inactivation of a Myoviridae bacteriophage infecting esters by immobilized lipase on magnetic Bacillus amyloliquefaciens FB11. Annals of microparticles. Bioscience Biotechnology and Microbiology, 65(4), 1841-1846. Biochemistry, 79(11), 1750-1758. Appendices 73
135. Prathumpai, W. and Kocharin, K. (2015). Phomalactone 142. Raita, M., Arnthong, J., Champreda, V. and optimization and production of entomopathogenic Laosiripojana, N. (2015). Modification of magnetic fungi by Ophiocordyceps communis BCC 1842 and BCC nanoparticle lipase designs for biodiesel production 2763. Preparative Biochemistry and Biotechnology, from palm oil. Fuel Processing Technology, 134, 189- 46(1), 44-48. 197.
136. Prompetchara, E., Ketloy, C., Keelapang, P., Sittisombut, 143. Raita, M., Kiatkittipong, W., Laosiripojana, N. and N. and Ruxrungtham, K. (2015). The immunogenicity Champreda, V. (2015). Kinetic study on esterification of tetravalent dengue DNA vaccine in mice pre- of palmitic acid catalyzed by glycine-based crosslinked exposed to Japanese encephalitis or Dengue virus protein coated microcrystalline lipase. Chemical antigens. Asian Pacific Journal of Allergy and Engineering Journal, 278, 19-23. Immunology, 33(3), 182-188. 144. Raita, M., Laosiripojana, N. and Champreda, V. (2015). 137. Pungrasmi, W., Chaisri, R., Malaphol, E. and Biocatalytic methanolysis activities of cross-linked Powtongsook, S. (2015). Efficiency of a hybrid solid protein-coated microcrystalline lipase toward digestion-denitrification column in suspended solid esterification/transesterification of relevant palm and nitrate removal from recirculating aquaculture products. Enzyme and Microbial Technology, 70, 28– system. Environmental Engineering Research, 20(2), 34. 175-180. 145. Ramli, R.A., Pudjiastuti, P., Tjahjandaric, T.S., Lie, W., 138. Punya, J., Swangmaneecharern, P., Pinsupa, S., Rattanajak, R., Kamchonwongapaisan, S. and Pyne, Nitistaporn, P., Ponghanphot, S., Kunathigan, V., S.G. (2015). Alkaloids from the roots of Stemona Cheevadhanarak, S., Tanticharoen, M. and javanica (Kunth) Engl. (Stemonaceae) and their anti- Amnuaykanjanasin, A. (2015). Phylogeny of type I malarial, acetylcholinesterase inhibitory and cytotoxic polyketide synthases (PKSs) in fungal entomopathogens activities. Phytochemistry Letters, 11, 157–162. and expression analysis of PKS genes in Beauveria 146. Regmi, S.M., Chaiprasert, A., Kulawonganunchai, S., bassiana BCC 2660. Fungal Biology, 119(6), 538–550. Tongsima, S., Coker, O.O., Prammananan, T., Viratyosin, F139. Punyadee, N., Mairiang, D., Thiemmeca, S., Komoltri, W. and Thaipisuttikul, I. (2015). Whole genome C., Pan-ngum, W., Chomanee, N., Charngkaew, K., sequence analysis of multidrug‑resistant Tangthawornchaikul, N., Limpitikul, W., Vasanawathana, Mycobacterium tuberculosis Beijing isolates from an S., Malasit, P. and Avirutnan, P. (2015). Microparticles outbreak in Thailand. Molecular Genetics and provide a novel biomarker to predict severe clinical Genomics, 290(5), 1933-1941. outcomes of dengue virus infection. Journal of 147. Regmi, S.M., Coker, O.O., Kulawonganunchai, S., Virology, 89(3), 1587-1607. Tongsima, S., Prammananan, T., Viratyosin, W., 140. Punyawai, K., Anakkul, N., Srirattana, K., Aikawa, Y., Thaipisuttikul, I. and Chaiprasert, A. (2015). Sangsritavong, S., Nagai, T., Imai, K. and Parnpai, R. Polymorphisms in drug-resistant-related genes shared (2015). Comparison of Cryotop and micro volume air among drug-resistant and pan-susceptible strains of cooling methods for cryopreservation of bovine sequence type 10, Beijing family of Mycobacterium matured oocytes and blastocysts. Journal of tuberculosis. International Journal of Mycobacteriology, Reproduction and Development, 61(5), 431-437. 4(1), 67-72.
141. Puseenam, A., Tanapongpipat, S. and Roongsawang, N. (2015). Co-expression of Endoxylanase and Endoglucanase in Scheffersomyces stipitis and Its Application in Ethanol Production.Applied Biochemistry and Biotechnology, 177(8), 1690-1700. 74 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
148. Réjou-Méchain, M., Muller-Landau, H. C., Detto, M., 154. Ruengphayak, S., Ruanjaichon, V., Saensuk, C., Thomas, S.C., Toan, T.L., Saatchi, S. S., Barreto-Silva, Phromphan, S., Tragoonrung, S., Kongkachuichai, R. J. S., Bourg, N. A., Bunyavejchewin, S., Butt, N., and Vanavichit, A. (2015). Forward screening for Brockelman, W. Y., Cao, M., Cárdenas, D., Chiang, seedling tolerance to Fe toxicity reveals a polymorphic J.-M., Chuyong, G.B., Clay, K., Condit, R., Dattaraja, H. mutation in ferric chelate reductase in rice.RICE , 8, 3. S., Davies, S. J., Duque, A., Esufali, S., Ewango, C., 155. Rukachaisirikul, V., Rungsaiwattana, N., Klaiklay, S., Fernando, R.H.S., Fletcher, C.D., Gunatilleke, I. A. U. Phongpaichit, S., Borwornwiriyapan, K. and Sakayaroj, N., Hao, Z., Harms, K. E., Hart, T. B., Hérault, B., Howe, J. (2014). γ-Butyrolactone, Cytochalasin, Cyclic R. W., Hubbell, S. P., Johnson, D. J., Kenfack, D., Larson, Carbonate, Eutypinic Acid, and Phenalenone A. J., Lin, L., Lin, Y., Lutz, J. A., Makana, J.-R., Malhi, Derivatives from the Soil Fungus Aspergillus sp. PSU- Y., Marthews, T. R., McEwan, R. W., McMahon, S. M., RSPG185. Journal of Natural Products, 77(11), 2375- McShea, W. J., Muscarella, R., Nathalang, A., Noor, N. 2382. S. M., Nytch,C. J., Oliveira, A. A., Phillips, R.P., Pongpattananurak, N., Punchi-Manage, R., Salim, R., 156. Rungrassamee, W., Kingcha, Y., Srimarut, Y., Schurman, J., Sukumar, R., Suresh, H. S., Suwanvecho, Maibunkaew, S., Karoonuthaisiri, N. and Visessanguan, U., Thomas, D. W., Thompson, J., Uríarte, M., Valencia, W. (2014). Mannooligosaccharides from copra meal R., Vicentini, A., Wol, A. T., Yap, S., Yuan,Z., Zartman, improves survival of the Pacific white shrimp C. E., Zimmerman, J. K., and Chave, J. (2014). Local (Litopenaeus vannamei) after exposure to Vibrio spatial structure of forest biomass and its consequences harveyi. Aquaculture, 434, 403–410. for remote sensing of carbon stocks. Biogeosciences Discuss, 11, 5711-5742. 157. Rungruengphol, C., Jaresitthikunchai, J., Wikan, N., Phaonakrop, N., Keadsanti, S., Yoksan, S., Roytrakul, 149. Ritcharoen, W., Powtongsook, S., Kangvansaichol, K. S. and Smith, D.R. (2015). Evidence of plasticity in the
and Pavasant, P. (2015). Effect of Daytime CO2 dengue virus: Host cell interaction. Microbial Supplement on Productivity and Biochemical Pathogenesis, 86, 18-25. Composition of Scenedesmus Armatus Under Outdoor Cultivation. Preparative Biochemistry and 158. Saengchan, K., Nopharatana, M., Lerdlattaporn, R. and Biotechnology, doi:10.1080/10826068.2015.101556 Songkasiri, W. (2015). Enhancement of starch-pulp 9. separation in centrifugal-filtration process: Effects of particle size and variety of cassava root on free starch 150. Ritcharoen, W., Sriouam, P., Nakseedee, P., Sang, P., granule separation.Food and Bioproducts Processing, Powtongsook, S., Kungvansaichol, K. and Pavasant, P. 95, 208-217. (2014). Cultivation options for indoor and outdoor growth of Chaetoceros gracilis with airlift 159. Saepua, S., Kornsakulkarn, J., Choowong, W., Supothina, photobioreactors. Maejo International Journal of S. and Thongpanchang, C. (2015). Bioxanthacenes and Science and Technology, 8(01), 100-113. monomeric analogues from insect pathogenic fungus Conoideocrella luteorostrata Zimm. BCC 31648. 151. Romyanon, K., Mosaleeyanon, K. and Kirdmanee, C. Tetrahedron, 71(16), 2400–2408. (2015). Direct-shoot organogenesis as an alternative protocol for in vitro regeneration of oil palm (Elaeis 160. Sakkhachornphop, S., Thongkum, W. and Tayapiwatana, guineensis Jacq.). Scientia Horticulturae, 195, 1-7. C. (2015). Novel 3′-Processing Integrase Activity Assay by Real-Time PCR for Screening and Identification of 152. Rucksaken, R., Haonon, O., Pinlaor, P., Pairojkul, C., HIV-1 Integrase Inhibitors. BioMed Research Roytrakul, S., Yongvanit, P., Selmi, C. and Pinlaor, S. International, 2015(2015), Aritcle ID 853891. (2015). Plasma IgG autoantibody against actin-related protein 3 in liver flukeOpisthorchis viverrini infection. 161. Sangsuriya, P., Charoensapsri, W., Chomwong, S., Parasite Immunology, 37(7), 340-348. Senapin, S., Tassanakajon, A. and Amparyup, P. (2015). A shrimp pacifastin light chain-like inhibitor: Molecular 153. Ruengphayak, S., Chaichumpoo, E., Phromphan, S., identification and role in the control of the Kamolsukyunyong, W., Sukhaket, W., Phuvanartnarubal, prophenoloxidase system. Developmental and E., Korinsak, S., Korinsak, S. and Vanavichit, A. (2015). Comparative Immunology, 54(1), 32-45. Pseudo-backcrossing design for rapidly pyramiding multiple traits into a preferential rice variety. RICE, 8, 7. Appendices 75
162. Sathasivam, R., Praiboon, J., Chirapart, A., 170. Simon-Loriere, E., Lin, R.J., Kalayanarooj, S.M., Trakulnaleamsai, S., Kermanee, P., Roytrakul, S. and Chuansumrit, A., Casademont, I., Lin, S.Y., Yu, H.P., Juntawong, N. (2014). Screening, phenotypic and Lert-itthiporn, W., Chaiyaratana, W., genotypic identification of β-carotene producing Tangthawornchaikul, N., Tangnararatchakit, K., strains of Dunaliella salina from Thailand. Indian Vasanawathana, S., Chang, B.L., Suriyaphol, P., Yoksan, Journal of Geo Marine Sciences, 43(12), 1-19. S., Malasit, P., Despres, P., Paul, R., Lin, Y.L. and Sakuntabhai, A. (2015). High Anti-Dengue Virus 163. Seepiban, C., Charoenvilaisiri, S., Kumpoosiri, M., Activity of the OAS Gene Family Is Associated With Bhunchoth, A., Chatchawankanphanich, O. and Increased Severity of Dengue. Journal of Infectious Gajanandana, O. (2015). Development of a protocol Diseases, 212(12), 2011-2020. for the identification of tospoviruses and thrips species in individual thrips. Journal of Virological Methods, 171. Sinsuwan, S., Jangchud, A., Rodtong, S., Roytrakul, S. 222, 206-213. and Yongsawatdigul, J. (2015). Statistical Optimization of the Production of NaCl-Tolerant Proteases by a 164. Sele, A.M., Bremner, J.B., Willis, A.C., Haritakun, R., Moderate Halophile, Virgibacillus sp. SK37. Food Griffith,R. and Keller, P.A. (2015). A cascade synthetic Technology and Biotechnology, 53(2), 136–145. route to new bioactive spiroindolinepyrido[1,2-a] indolediones from Indirubin. Tetrahedron, 71(43), 172. Sinthujaroena, P., Tonganunt-Srithawornb, M., 8357–8367. Eurwilaichitrc, L. and Phongdara, A. (2015). Protection of Litopenaeus vannamei against the white spot 165. Senachak, J., Cheevadhanarak, S. and Hongsthong, A. syndrome virus using recombinant Pm-fortilin (2015). SpirPro: A Spirulina proteome database and expressed in Pichia pastoris. Aquaculture, 435, 450- web-based tools for the analysis of protein-protein 457. interactions at the metabolic level in Spirulina (Arthrospira) platensis C1. BMC Bioinformatics, 16, 173. Sirikharin, R., Taengchaiyaphum, S., Sanguanrut, P., 233. Chi, T.D., Mavichak, R., Proespraiwong, P., Nuangsaeng, B., Thitamadee, S., Flegel, T.W. and Sritunyalucksana, 166. Sengprasert, P., Amparyup, P., Tassanakajorn, A. and K. (2015). Characterization and PCR Detection Of F Wongpanya, R. (2015). Characterization and Binary, Pir-Like Toxins from Vibrio parahaemolyticus identification of calmodulin and calmodulin binding Isolates that Cause Acute Hepatopancreatic Necrosis proteins in hemocyte of the black tiger shrimp Disease (AHPND) in Shrimp. PLOS ONE, 10(5), (Penaeus monodon). Developmental and Comparative e0126987. Immunology, 50(2), 87–97. 174. Siripipatthana, P., Phaonakrop, N., Roytrakul, S., 167. Sereekhajornjaru, N., Somboon, C., Rattanajak, R., Senawong, G., Mudalige-Jayawickrama, R.G. and Denny, W.A., Wilairat, P. and Auparakkitanon, S. (2014). Sattayasai, N. (2015). The first trimeric Galanthus Comparison of hematin-targeting properties of nivalis agglutinin-related lectin of Orchidaceae was pynacrine, an acridine analog of the benzonaphthyridine found in Dendrobium pendulum: purification, antimalarial pyronaridine.Acta Tropica, 140, 181-183. characterization, and effects of stress factors. Plant 168. Shakoori, A., Bremner, J.B., Abdel-Hamid, M.K., Willis, Cell Reports, 34(7), 1253-1262. A.C., Haritakun, R. and Keller, P.A. (2015). Further 175. Somyong, S., Poopear, S., Jomchai, N., Uthaipaisanwong, exploration of the heterocyclic diversity accessible P., Ruang-areerate, P., Sangsrakru, D., Sonthirod, C., from the allylation chemistry of indigo. Beilstein Ukoskit, K., Tragoonrung, S. and Tangphatsornruang, Journal of Organic Chemistry, 11, 481–492. S. (2015). The AKR gene family and modifying sex ratios 169. Shearman, J.R., Sangsrakru, D., Jomchai, N., Ruang- in palms through abiotic stress responsiveness. Areerate, P., Sonthirod, C., Naktang, C., Functional and Integrative Genomics, 15(3), 349-362. Theerawattanasuk, K., Tragoonrung, S. and 176. Soonthornchai, W., Chaiyapechara, S., Jarayabhand, Tangphatsornruang, S. (2015). SNP Identification from P., Söderhäll, K. and Jiravanichpaisal, P. (2015). RNA Sequencing and Linkage Map Construction of Interaction ofVibrio spp. with the Inner Surface of the Rubber Tree for Anchoring the Draft Genome. PLOS Digestive Tract ofPenaeus monodon. PLOS ONE, 10(8), ONE, 10(4), e0121961. e0135783. 76 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
177. Soowannayan, C., Nguyen, G.T., Pham, L.N., Phanthura, 184. Suphatrakul, A., Yasanga, T., Keelapang, P., Sriburi, R., M. and Nakthong, N. (2015). Australian red claw Roytrakul, T., Pulmanausahakul, R., Utaipat, U., crayfish (Cherax quadricarinatus) is susceptible to Kawilapan, Y., Puttikhunt, C., Kasinrerk, W., Yoksan, S., yellow head virus (YHV) infection and can transmit it Auewarakul, P., Malasit, P., Charoensri, N. and to black tiger shrimp (Penaeus monodon). Aquaculture, Sittisombut, N. (2015). Generation and preclinical 445(1), 63–69. immunogenicity study of dengue type 2 virus-like particles derived from stably transfected mosquito 178. Sornwatana, T., Bangphoomi, K., Roytrakul, S., cells. Vaccine, 33(42), 5613-5622. Wetprasit, N., Choowongkomon, K. and Ratanapo, S. (2015). Chebulin: Terminalia chebula Retz. fruit- 185. Suraprasit, S., Methatham, T., Jaree, P., Phiwsaiya, K., derived peptide with angiotensin-I-converting enzyme Senapin, S., Hirono, I., Lo, C.F., Tassanakajon, A. and inhibitory activity. Biotechnology and Applied Somboonwiwat, K. (2014). Anti-lipopolysaccharide Biochemistry, 62(6), 746-753. factor isoform 3 from Penaeus monodon (ALFPm3) exhibits antiviral activity by interacting with WSSV 179. Sri-indrasutdhi, V., Tsui, C.K.M., Chuaseeharonnachai, structural proteins. Antiviral Research, 110, 142–150. C., Yamaguchi, K., Suetrong, S., Okane, I., Nakagiri, A. and Boonyuen, N. (2015). Helicocentralis hyalina gen. 186. Suriyachadkun, C., Ngaemthao, W., Chunhametha, et sp. nov., an aero-aquatic helicosporous fungus S. and Sanglier, J.J. (2015). Actinoplanes luteus sp. nov., (Leotiomycetes, Ascomycota) in Thailand.Mycological isolated from soil in Thailand. International Journal of Progress, 14, 81. Systematic and Evolutionary Microbiology, doi: 10.1099/ijsem.0.000562. 180. Srilohasin, P., Chaiprasert, A., Tokunaga, K., Nishida, N., Prammananan, T., Smittipat, N., Mahasirimongkol, 187. Suriyachadkun, C., Ngaemthao, W., Chunhametha, S., S., Chaiyasirinroje, B., Yanai, H. and Palittapongarnpim, Thawai, C. and Sanglier, J.J. (2015). Sinosporangium P. (2014). Genetic Diversity and Dynamic Distribution fuscum sp. nov., isolated from soil in Thailand. of Mycobacterium tuberculosis Isolates Causing International Journal of Systematic and Evolutionary Pulmonary and Extrapulmonary Tuberculosis in Microbiology, 65(6), 435–444. Thailand. Journal of Clinical Microbiology, 52(12), 188. Surussawadee, J., Jindamorakot, S., Nakase, T., Lee, 4267-4274. CF. and Limtong, S. (2015). Hannaella phyllophila sp. 181. Srisucharitpanit, K., Yao, M., Promdonkoy, B., nov., a basidiomycetous yeast species associated with Chimnaronk, S., Tanak, I. and Boonserm, P. (2014). plants in Thailand and Taiwan. International Journal Crystal structure of BinB: A receptor binding component of Systematic and Evolutionary Microbiology, 65(7), of the binary toxin from Lysinibacillus sphaericus. 2135-2140. Proteins-Structure Function and Bioinformatics, 189. Sutthangkul, J., Amparyup, P., Charoensapsri, W., 82(10), 2703–2712. Senapin, S., Phiwsaiya, K. and Tassanakajon, A. (2015). 182. Sri-Uam, P., Linthong, C., Powtongsook, S., Suppression of Shrimp Melanization during White Spot Kungvansaichol, K. and Pavasant, P. (2015). Syndrome Virus Infection. Journal of Biological Manipulation of Biochemical Compositions ofChlorella Chemistry, 290(10), 6470-6481. sp.. Engineering Journal, 19(4), 13-24. 190. Swetwiwathana, A. and Visessanguan, W. (2015). 183. Suebsing, R., Kampeera, J., Sirithammajak, S., Potential of bacteriocin-producing lactic acid bacteria Withyachumnarnkul, B., Turner, W. and Kiatpathomchai, for safety improvements of traditional Thai fermented W. (2015). Colorimetric Method of Loop-Mediated meat and human health. Meat Science, 109, 101–105. Isothermal Amplification with the Pre-Addition of 191. Tadpetch, K., Chukong, C., Jeanmard, L, Thiraporn, A., Calcein for Detecting Flavobacterium columnare and Rukachaisirikul, V., Phongpaichit, S. and Sakayaroj, J. its Assessment in Tilapia Farms. Journal of Aquatic (2015). Cytotoxic naphthoquinone and a new succinate Animal Health, 27(1), 38-44. ester from the soil fungus Fusarium solani PSU- RSPG227. Phytochemistry Letters, 11, 106–110. Appendices 77
192. Talawanicha, Y., Kamchonwongpaisana, S., 200. Thongkum, M., Burns, P., Bhunchoth, A., Warin, N., Sirawarapornb, W. and Yuthavong, Y. (2015). Use of Chatchawankanphanich, O. and Doorn, W.G.V. (2015). bacterial surrogates as a tool to explore antimalarial Ethylene and pollination decrease transcript drug interaction: Synergism between inhibitors of abundance of an ethylene receptor gene in Dendrobium malarial dihydrofolate reductase and dihydropteroate petals. Journal of Plant Physiology, 176, 96-100. synthase. Acta Tropica, 149, 64–69. 201. Tong-Ngam, P., Roytrakul, S. and Sritanaudomchai, H. 193. Tangsongcharoen, C., Chomanee, N., Promdonkoy, B. (2015). BmKn-2 Scorpion Venom Peptide for Killing and Boonserm, P. (2015). Lysinibacillus sphaericus Oral Cancer Cells by Apoptosis. Asian Pacific Journal binary toxin induces apoptosis in susceptible Culex of Cancer Prevention, 16(7), 2807-2811. quinquefasciatus larvae. Journal of Invertebrate 202. Toopcham, T., Roytrakul, S. and Yongsawatdigul, J. Pathology, 128, 57–63. (2015). Characterization and identification of 194. Thammasit, P., Sangboonruang, S., Suwanpairoj, S., angiotensin I-converting enzyme (ACE) inhibitory Khamaikawin, W., Intasai, N., Kasinrerk, W., peptides derived from tilapia using Virgibacillus Tayapiwatana, C. and Tragoolpua, K. (2015). Intracellular halodenitrificans SK1-3-7 proteinases. Journal of Acidosis Promotes Mitochondrial Apoptosis Pathway: Functional Foods, 14, 435–444. Role of EMMPRIN Down-regulationvia Specific Single- 203. Trana, T., Da, G., Moreno-Santander, M.A., Vélez- chain Fv intrabody. Journal of Cancer, 6(3), 276-286. Hernández, G.A., Giraldo-Toro, A., Piyachomkwan, K., 195. Tharad, S., Iturri, J., Moreno-Cencerrado, A., Sriroth, K. and Dufour, D. (2015). A comparison of Mittendorfer, M., Promdonkoy, B., Krittanai, CK. and energy use, water use and carbon footprint of cassava Toca-Herrera, J.L. (2015). Effect of the Concentration starch production in Thailand, Vietnam and Colombia. of Cytolytic Protein Cyt2Aa2 on the Binding Mechanism Resources Conservation and Recycling, 100, 31–40. on Lipid Bilayers Studied by QCM-D and AFM. 204. Tsutsumi, K., Yamada, N., Cha-um, S., Tanaka, Y. and Langmuir, 31(38), 10477-10483. Takabe, T. (2015). Differential accumulation of 196. Theerawitaya, C., Tisarum, R., Samphumphuang, T., glycinebetaine and choline monooxygenase in bladder F Singh, H.P., Cha-Um, S., Kirdmanee, C. and Takabe, T. hairs and lamina leaves of Atriplex gmelini under high (2015). Physio-biochemical and morphological salinity. Journal of Plant Physiology, 176, 101-107. characters of halophyte legume shrub, Acacia 205. Uawisetwathana, U., Graham, S.F., Kamolsukyunyong, ampliceps seedlings in response to salt stress under W., Sukhaket, W., Klanchui, A., Toojinda, T., Vanavichit, greenhouse. Frontiers in Plant Science, 6, 630. A., Karoonuthaisiri, N. and Elliott, C.T. (2015). 197. Theerawitaya, C., Yamada, N., Samphumphuang, T., Quantitative 1H NMR metabolome profiling of Thai Cha-um, S., Kirdmanee, C. and Takabe, T. (2015). Jasmine rice (Oryza sativa) reveals primary metabolic Evaluation of Na+ enrichment and expression of some response during brown planthopper infestation. carbohydrate related genes in indica rice seedlings Metabolomics, doi:10.1007/s11306-015-0817-4. under salt stress. Plant Omics, 8(2), 130-140. 206. Unrean, P. and Franzen, C.J. (2015). Dynamic flux 198. Thipubol, S., Uthaipibull, C., Kamchonwongpaisan, S., balancing elucidates NAD(P)H production as limiting Tipsuwan, W. and Srichairatanakool, S. (2015). response to furfural inhibition in Saccharomyces Inhibitory effect of novel iron chelator, 1-(N-acetyl-6- cerevisiae. Biotechnology Journal, 10(8), 1248-1258. aminohexyl)-3-hydroxy-2-methylpyridin-4-one (CM1) 207. Urtgam, S., Treerattrakool, S., Roytrakul, S., Wongtripop, and green tea extract on growth of Plasmodium S., Prommoon, J., Panyim, S. and Udomkit, A. (2015). falciparum. Malaria Journal, 14, 382. Correlation between gonad-inhibiting hormone and 199. Thipubon, P., Tipsuwan, W., Uthaipibull, C., vitellogenin during ovarian maturation in the Santitherakul, S. and Srichiratanakool, S. (2015). Anti- domesticated Penaeus monodon. Aquaculture, 437, malarial effect of 1-(N-acetyl-6-aminohexyl)-3- 1-9. hydroxy-2-methylpyridin-4-one and green tea extract on erythrocyte-stage Plasmodium berghei in mice. Asian Pacific Journal of Tropical Biomedicine, 5(11), 932-936. 78 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
208. Visetnan, S., Donpudsa, S., Supungul, P., Tassanakajon, 216. Whangchenchom, W., Chiemchaisri, W., A. and Rimphanitchayakit, V. (2014). Domain 2 of a Tapaneeyaworawong, P. and Powtongsook, S. (2014). Kazal serine proteinase inhibitor SPIPm2 from Penaeus Wastewater from Instant Noodle Factory as the Whole monodon possesses antiviral activity against WSSV. Nutrients Source for the Microalga Scenedesmus sp. Fish and Shellfish Immunology, 41(2), 526-530. Cultivation. Environmental Engineering Research, 19(3), 283-287. 209. Visetnan, S., Supungul, P., Hirono, I., Tassanakajon, A. and Rimphanitchayakit, V. (2015). Activation of 217. Whangchenchom, W., Chiemchaisri, W., PmRelish from Penaeus monodon by yellow head virus. Tapaneeyaworawong, P. and Powtongsook, S. (2015). Fish and Shellfish Immunology, 42(2), 335-344. Production of microalgal biomass using raw wastewater from instant noodle factory. The International Journal 210. Vorapreeda, T., Thammarongtham, C., Cheevadhanarak, of Current Research and Academic Review, Special S. and Laoteng, K. (2015). Genome mining of fungal Issue-2, 190-199. lipid-degrading enzymes for industrial applications. Microbiology, 161(8), 1613-1626. 218. Wijayawardene, N.N., Crous, P.W., Kirk, P.M., Hawksworth, D.L., Boonmee, S., Braun, U., Dai, D.Q., 211. Vu, H.T.L., Malimas, T., Chaipitakchonlatarn, W., Bui, D’souza, M.J., Diederich, P., Dissanayake, A., Doilom, V.T.T., Yukphan, P., Bui, U.T.T., Muramatsu, Y., Sitdhipol, M., Hongsanan, S., Jones, E.B.G., Groenewald, J.Z., J., Tanasupawat, S., Duong, K.C., Nakagawa, Y., Pham, Jayawardena, R., Lawrey, J.D., Liu, J.K., Lücking, R., H.T. and Yamada, Y. (2015). Tanticharoenia aidae sp. Madrid, H., Manamgoda, D.S., Muggia, L., Nelsen, M.P., nov., for acetic acid bacteria isolated in Vietnam. Phookamsak, R., Suetrong, S., Tanaka, K., Thambugala, Annals of Microbiology, doi:10.1007/s13213-015- K.M., Wanasinghe, D.N., Wikee, S., Zhang, Y., Aptroot, 1124-z. A., Ariyawansa, H.A., Bahkali, A.H., Bhat, D.J., Gueidan, 212. Waiwijit, U., Phokaratkul, D., Kampeera, J., Lomas, T., C., Chomnunti, P., Hoog, G.H.D., Knudsen, K., Li, W.J., Wisitsoraat, A., Kiatpathomchai, W. and Tuantranont, McKenzie, E.H.C., Miller, A.N., Phillips, A.J.L., Piątek, A. (inpress). Graphene oxide based fluorescence M., Raja, H.A., Shivas, R.S., Slippers, B., Taylor, J.D., resonance energy transfer and loop-mediated Tian, Q., Wang, Y., Woudenberg, J.H.C., Cai, L., Jaklitsch, isothermal amplification for white spot syndrome virus W.M. and Hyde, K.D. (2014). Naming and outline of detection. Journal of Biotechnology, 212, 44-49. Dothideomycetes–2014 including proposals for the protection or suppression of generic names. Fungal 213. Warit, S., Phunpruch, S., Jityam, C., Jaitrong, S., Diversity, 69(1), 1-55. Billamas, P., Chaiprasert, A., Palittapongarnpim, P. and Prammananan, T. (2015). Genetic characterisation of 219. Wintachai, P., Kaur, P., Lee, R.C.H., Ramphan, S., a whiB7 mutant of a Mycobacterium tuberculosis Kuadkitkan, A., Wikan, N., Ubol, S., Roytrakul, S., Chu, clinical strain. Journal of Global Antimicrobial J. J. H. and Smith, D.R. (2015). Activity of andrographolide Resistance, 3(4), 262-266. against chikungunya virus infection.Scientific Reports, 5, 14179. 214. Wasuworawong, K., Roytrakul, S., Paemanee, A., Jindapornprasert, K. and Komyod, W. (2015). 220. Wintachai, P., Thuaud, F., Basmadjian, C., Roytrakul, Comparative Proteomic Analysis of Human S., Ubol, S., Désaubry, L. and Smith, D.R. (2015). Cholangiocarcinoma Cell Lines: S100A2 as a Potential Assessment of flavaglines as potential chikungunya Candidate Protein Inducer of Invasion. Disease virus entry inhibitors. Microbiology and Immunology, Markers, Article ID 629367. 59(3), 129-141.
215. Wattanakulpakin, T., Iamtham, S., Grubbs, K.C. and Volkaert, H.A. (2015). Population genetic analysis of Xylia xylocarpa (Fabaceae—Mimosoideae) in Thailand. Tree Genetics and Genomes, 11, 7. Appendices 79
221. Witschel, M.C., Rottmann, M., Schwab, A., 227. Xie, X., Zou, J., Puttikhunt, C., Yuan, Z. and Shi, P.Y. Leartsakulpanich, U., Chitnumsub, P., Seet, M., Tonazzi, (2015). Two distinct sets of NS2A molecules are S., Schwertz, G., Stelzer, F., Mietzner, T., McNamara, responsible for dengue virus RNA synthesis and virion C., Thater, F., Freymond, C., Jaruwat, A., Pinthong, C., assembly. Journal of Virology, 89(2), 1298-1313. Riangrungroj, P., Oufir, M., Hamburger, M., Mäser, P., 228. Yamada, N., Theerawitaya, C., Kageyama, H., Cha-um, Sanz-Alonso, L.M., Charman, S., Wittlin, S., Yuthavong, S. and Takabe, T. (2015). Expression of developmentally Y., Chaiyen, P. and Diederich, F. (2015). Inhibitors of regulated plasma membrane polypeptide (DREPP2) Plasmodial Serine Hydroxymethyltransferase (SHMT): in rice root tip and interaction with 2+Ca /CaM complex Cocrystal Structures of Pyrazolopyrans with Potent and microtubule. Protoplasma, 252(6), 1519-1527. Blood- and Liver-Stage Activities.Journal of Medicinal Chemistry, 58(7), 3117–3130. 229. Yasamut, U., Yenchitsomanus, P., Junking, M., Noisakran, S., Puttikhunt, C., Hann-Chu, J.J. and 222. Wongratpanya, K., Imjongjairak, S., Waeonukul, R., Limjindaporn, T. (2015). Adaptor protein 1A facilitates Sornyotha, S., Phitsuwan, P., Pason, P., Nimchua, T., dengue virus replication.PLOS ONE, 10(6), e0130065. Tachaapaikoon, C. and Ratanakhanokchai, K. (2015). Multifunctional Properties of Glycoside Hydrolase 230. Yindeeyoungyeon, W., Rukseree, K., Tungsudjai, S. and Family 43 from Paenibacillus curdlanolyticus Strain Panichakul, T. (2015). Evaluation of Recombinant B-6 Including Exo-β-xylosidase, Endo-xylanase, and Mycobacterium tuberculosis Antigens MPT64, CFP10, α-L-Arabinofuranosidase Activities. Bioresources, and ESAT6 for Delayed-Type Hypersensitivity Responses 10(2), 2492-2505. in Guinea Pigs. Advances in Microbiology, 5(8), 586- 598. 223. Wongsombat, C., Aroonsri, A., Kamchonwongpaisan, S., Morgan, H.P., Walkinshaw, M.D., Yuthavong, Y. and 231. Yoiprommarat, S., Srichomthong, K., Deelai, S., Shaw, P.J. (2014). Molecular characterization of Suetrong, S., Sakayaroj, J., Bunyapaiboonsri, T. and Plasmodium falciparum Bruno/CELF RNA binding Unagul, P. (2015). Secondary metabolites of the marine proteins. Molecular and Biochemical Parasitology, fungus Paradendryphiellaarenariae BCC 17999. F 198(1), 1-10. Botanica Marina, 58(5), 393-399. 224. Wongtrakul, J., Thongtan, T., Roytrakul, S., Kumrapich, 232. Youngcharoen, S., Senapin, S., Lertwimol, T., Longyant, B., Janphen, K., Praparattanapan, J., Supparatpinyo, S., Sithigorngul, P., Flegel, T.W. and Chaivisuthangkura, K. and Smith, D.R. (2014). Proteomic Analysis of Serum P. (2015). Interaction study of a novelMacrobrachium and Urine of HIV-Monoinfected and HIV/HCV- rosenbergii effector caspase with B2 and capsid Coinfected Patients Undergoing Long Term Treatment proteins of M.rosenbergii nodavirus reveals their roles with Nevirapine. Disease Markers, Article ID 315824. in apoptosis. Fish and Shellfish Immunology, 45(2), 534-542. 225. Woradulayapinij, W., Audtho, M. and Wiwat, C. (2015). Suppression of α-glucosidase gene by RNA interference 233. Yundaeng, C., Somta, P., Tangphatsornruang, S., can increase resistance of Culex quinquefasciatus to Chankaew, S. and Srinives, P. (2015). A single base Bacillus sphaericus binary toxin. Biocontrol Science substitution in BADH /AMADH is responsible for and Technology, 25(12), 1453-1466. fragrance in cucumber (Cucumis sativus L.), and development of SNAP markers for the fragrance. 226. Woraprayote, W., Pumpuang, L., Tosukhowong, A., Theoretical and Applied Genetics, 128(9), 1881-1892. Roytrakul, S., Perez, R.H., Zendo, T., Sonomoto, K., Benjakul, S. and Visessanguan, W. (2015). Two putatively novel bacteriocins active against Gram- negative food borne pathogens produced byWeissella hellenica BCC 7293. Food Control, 55, 176–184. 80 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
List of Intellectual Properties List of Issued Intellectual Properties
Title Granted Date Ref. No. Country Patent Electrochemical detection of capsaicinoid 3 February 2015 US 8,945,370 B2 USA compounds in a sample The production process and water supporting 10 July 2015 44975 Thailand material made from cassava starch for tissue and cell culture Petty Patent Method to screen and classify Plasmodium 7 November 2014 9297 Thailand falciparum in blood sample Method to screen and classify Plasmodium vivax 7 November 2014 9298 Thailand in blood sample Specific primers for shrimp parasite,Enterocytozoon 29 December 2014 9405 Thailand hepatopenaei, and its application Method to induce mutation in plant using the 30 January 2015 9488 Thailand combination of controlled atmospheric pressure and mutagen Production process of recombinant lignocellulolytic 6 February 2015 9509 Thailand enzymes by a slow methanol utilization strain of Pichia pastoris Simulatneous desizing and scouring process for 10 April 2015 9768 Thailand natural fiber fabric using multi-activity enzymes Culture medium for growth of fungi isolated from 21 May 2015 9900 Thailand marine and mangrove habitats One-step desizing and scouring process of natural 21 May 2015 9901 Thailand fiber fabric using multi-enzyme The use of yeast cell with phytase on cell surface in 21 May 2015 9903 Thailand combination with a polysaccharide-degrading enzyme to improve nutrient in aninal feed Pulp bleaching process using alkaline-tolerant 10 July 2015 10069 Thailand xylanase obtained from metagenomic library of sugarcane bagasse compost Method for Listeria monocytogenes detection based 10 July 2015 10090 Thailand on antibody array using liposome encapsulation conjugated with antibodies and enzyme for naked eye signal detection Production process for enzyme - resistant rice 3 August 2015 10155 Thailand starch type III Detection of infectious myonecrosis virus using 7 August 2015 10190 Thailand immunochromatographic strip test Appendices 81
Title Granted Date Ref. No. Country Method to minimize released water and weight 7 August 2015 10191 Thailand losses in fermented meat products under acidic condition using heat-modified whey protein isolates Inoculum preparation of methanol utilization slow 7 August 2015 10192 Thailand (MutS) Pichia pastoris to improve fermentation efficiency Culture media to increase sporulation of blast 14 August 2015 10236 Thailand pathogen Method to induce mutation in plant tissue culture 28 August 2015 10305 Thailand under controlled environment Colorimetric detection method of methylparathion 28 August 2015 10306 Thailand based on the interaction of gold nanoparticle and methylparathion hydrolase Method to induce mutation in plant using the 28 August 2015 10308 Thailand combination of ultrasonic energy and mutagen Breeding flood-tolerant rice by mutation 28 August 2015 10309 Thailand Method to induce mutation in plant 28 August 2015 10310 Thailand DNA vector for expressing gene responsible for the 28 August 2015 10311 Thailand production of protein and metabolite in filamentous fungi Fermentation process of cassava feedstock with high 28 August 2015 10312 Thailand solid content for the production of biofuels and Fchemicals Detection of white spot syndrome virus, WSSV, with 4 September 2015 10337 Thailand LAMP-LFD technique Non-autoclave tissue culture media 4 September 2015 10339 Thailand Enzymatic process for pulp grinding in the paper- 4 September 2015 10340 Thailand making process Immunoassay based on flocculation of graphene 11 September 2015 10370 Thailand oxide particles Method to increase sporulation of rice-blast 18 September 2015 10412 Thailand pathogenic fungus (Magnaporthe grisea) using organic solvents to extract metabolites from blast- infected leaves Immunochromatographic strip test for multiplex 18 September 2015 10414 Thailand detection of three plant pathogens in cucurbits Plasmid vector for gene cloning in bacteria 18 September 2015 10415 Thailand 82 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
List of Applied Intellectual Properties
Title Filing Date Ref. No. Country Patent Cyclic peptide with broad spectrum inhibitions to 22 September 2014 1401005578 Thailand human pathogens Virus-like particle for the induction of immune 15 October 2014 1401006463 Thailand response against dengue virus infection and a method of generation Specific primers and DNA probes for SNP biomarkers 14 November 2014 1401006833 Thailand associated with the height of oil-palm trunk, and their applications Dengue serotype 2 virus-based plasmids for the 28 November 2014 1401007120 Thailand generation of codon-optimized, chimeric live attenuated vaccine candidates and the resultant viruses Method to quantify the contamination of aflatoxin- 23 December 2014 1401007690 Thailand producing mold Bioreagent for one-step desizing and biostoning 29 December 2014 1401007873 Thailand process of denim fabric Colloidal solution for acetone gas detection 13 February 2015 1501000778 Thailand Recombinant plasmid and recombinant Aspergillus 6 March 2015 1501001230 Thailand sp. for the production of unsaturated fatty acid, dihomo-γ-linolenic acid Process and formulation of non-sodium and non- 12 March 2015 1501001462 Thailand phosphate compounds for improving yield and textural quality of cooked chicken meat The process for the fabrication of polysalophen 26 May 2015 1501002871 Thailand complex on an electrode for the electrochemical determination of artemsinin and products for a method thereof Improving extraction yield in cassava starch factory 29 May 2015 1501002971 Thailand by recovery of starch in pulp at high solid loading with enzyme cocktails as a continuous integrated process Process of making cassava flour with high-viscosity 29 May 2015 1501002972 Thailand for small scale production Appendices 83
Title Filing Date Ref. No. Country Apparatus and method to enhance the emission of 4 June 2015 1501003071 Thailand multicolor optical molecules Anti-malarial and anti-cancer Phomoxydiene A 4 June 2015 1501003072 Thailand Steffimycin C, the potent antimycobacterial and 12 June 2015 1501003280 Thailand antiplasmodial agent Screen printed electrode with surface modification 10 July 2015 1501003963 Thailand for the determination of progesterone and the method thereof Enzyme formulation for converting lignocellulosic 29 July 2015 1501004284 Thailand biomass to sugar Using image processing to automatically identify 3 August 2015 1501004344 Thailand bands from gel electrophoresis image Detection method for antifolate-resistant malaria 28 August 2015 1501004966 Thailand pathogen Culture media process for production of treholose 28 August 2015 1501004968 Thailand by fermentation of thermotolerant yeast strain Method of preparation of bacterial cellulose/ 11 September 2015 1501005314 Thailand recombinant staterin-fibronectin fusion protein / calcium phosphate composites A method for genome-wide to identification of 11 September 2015 1501005315 Thailand transcription start sites in eukaryotes from NGS transcriptome data FCrop improvement using colchicine and sodium 18 September 2015 1501005652 Thailand azide by controlling light exposure-time Derivatives of 2,4-diamino-6-ethylpyrimidine 23 September 2015 1501005737 Thailand inhibiting Plasmodium falciparum Method of preparation of cellulose materials for 25 September 2015 1501005835 Thailand cartilage tissue engineering using reductive amination reduction in acidic conditions Protocol for isolation of live Acidovorax avenae 30 September 2015 1501006000 Thailand subsp. citrulli from cucumber and watermelon using immunomagnetic separation in combination with selective medium Gene expression system for the production of target 30 September 2015 1501006030 Thailand protein on yeast cell surface Petty Patent Preparation method of cassava starch-based 21 November 2014 1403001535 Thailand hydrogel for use as disintegrant in drug tablet Recombinant yeast Pichia stipitis directly produces 13 January 2015 1503000026 Thailand ethanol from cellulose, and its application Recombinant yeast Pichia stipitis directly produces 13 January 2015 1503000027 Thailand ethanol from cellulose and hemicellulose, and its application 84 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Title Filing Date Ref. No. Country Detection method for Streptococcus agalactiae in 27 March 2015 1503000427 Thailand fish Detection method for Streptococcus iniae in fish 27 March 2015 1503000428 Thailand Detection method forAeromonas hydrophila in fish 27 March 2015 1503000429 Thailand Detection method for Francisella noatuensis subsp. 27 March 2015 1503000430 Thailand orientalis in fish Detection method for Infectious Spleen and Kidney 27 March 2015 1503000431 Thailand Necrosis Virus in fish Screening method for latent infection in sugarcane 10 April 2015 1503000530 Thailand stalk Method to improve crop yield using the combination 10 April 2015 1503000531 Thailand of high carbon dioxide treatment and mutagen Method to improve salinity and drought tolerance 10 April 2015 1503000532 Thailand in soybean using the combination of plant growth regulator and mutagen Recombinant plasmid for extracellular expression 10 April 2015 1503000867 Thailand of heterologous protein in yeast Ogataea spp. Method to improve crop yield in rice using the 19 June 2015 1503000930 Thailand combination of high concentration sugar syrup, mutagen and utltrasonic energy Test kit for multi-drug resistant tuberculosis using 14 August 2015 1503001228 Thailand nucleic acid lateral flow immunochromatography Method of preparing nanocomposite material from 19 August 2015 1503001250 Thailand magnetic nanoparticles and cationic starch for chromium (VI) absorption Culture media and cultivation process to enhance 19 August 2015 1503001251 Thailand mycelium growth of Boletus mushroom Bioreagent for desizing and scouring process of 4 September 2015 1503001399 Thailand fabric and its application The process of plant improvement for environmental 4 September 2015 1503001400 Thailand stress conditions using direct current and mutagen Method of preparation of porous cellulose materials 25 September 2015 1503001571 Thailand for cell support A marker recycling DNA tool for genome modification 25 September 2015 1503001578 Thailand of yeast strain Ogataea thermomethanolica and its application Appendices 85
Title Filing Date Ref. No. Country Immunomagnetic beads for specific isolation of 30 September 2015 1503001617 Thailand Acidovorax avenae subsp. citrulli Monoclonal antibody to nucleocapsid protein of 30 September 2015 1503001618 Thailand Tomato necrotic ringspot virus (TNRV) found in Thailand and its application in immunodetection of TNRV Automatic thalassemia interpretation and couple 30 September 2015 1503001623 Thailand at-risk assessment system for accurately identifying types of thalassemia from hemoglobin typing profiles Trade Secret Solid cultured formulation for the preparation of 1 March 2015 N/A Thailand A. niger BCC5639 inoculum Method for multi-enzyme production fromA. niger 1 March 2015 N/A Thailand BCC5639
F 86 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Honors and Awards
Dr. Noppol Kobmoo Dr. Bunpote Siridechadilok Bioresources Technology Research Unit Medical Biotechnology Research Laboratory Marie Skłodowska-Curie Individual Fellowship to The 2014 Young Technologist Award in recognition work with Prof. Titiana Giraud at the Ecology, of the development of virus construction that can Systematics and Evolution Laboratory of the save time and effort and enables the application University of Paris-Sud, France on a project entitled of diverse genetic approaches, awarded by the “Insights from Population Genomics to the Evolution Foundation for the Promotion of Science and of Host Specificity in Insect Fungi (GenoSpec)” for Technology under the Patronage of His Majesty the 2 years starting from November 2015 to 2017, King. awarded by Horizon 2020, the EU Framework Programme for Research and Innovation. Dr. Chalermpol Kirdmanee Genome Technology Research Unit Dr. Atikorn Panya SCG Chair Professor Scholarship in recognition of Food Biotechnology Research Unit his work on saline land rehabilitation, awarded by In collaboration with King Mongkut’s University of Siam Cement Group. Technology North Bangkok and the University of Massachusetts Amherst (USA) Ms. Wansika Kiatpathomchai, Dr. Rungkarn Suebsing The 2015 Edwin Frankel Award for Best Paper in and Ms. Jantana Kampeera Lipid Oxidation and Quality for the paper titled Biosensing Technology Research Unit “Impact of free fatty acids and phospholipids on In collaboration with the National Center for Electronics reverse micelles formation and lipid oxidation in and Computer Technology Center bulk oil”, selected by the American Oil Chemists’ The 2015 NRCT Invention Award (Agricultural Society, (AOCS) to recognize the best paper relating Science and Biology) for “Portable rapid and low to lipid oxidation or quality published during the cost aflatoxin sensor with LAMP-Electrochemical past year in AOCS Press publications. detection kit”, awarded by the National Research Council of Thailand. Oil palm breeding and the development of tenera palm regeneration system through somatic Dr. Nitsara Karoonuthaisiri, Dr. Rungnapa Leelatanawit, embryogenesis Ms. Umaporn Uawisetwathana, Ms. Thidathip The 2015 Outstanding Project to recognize projects Wongsurawat, Ms. Amornpan Klanchui and that have generated socio-economic impact, Ms. Natechanok Thamniemdee awarded by Agricultural Research Development Biosensing Technology Research Unit and Animal Agency (Public Organization). Biotechnology Research Unit The 2014 NRCT Research Award (Agricultural Dr. Theerayut Toojinda Science and Biology Field) for “The Development Rice Gene Discovery Laboratory of DNA Microarray Technology to Address Poor The 2015 Plant Breeder Award in recognition of his Reproductive Maturation Problems in the Black success in utilizing marker-selected selection Tiger Shrimp (Peneaus monodon)”, awarded by the technique in rice improvement, presented by the National Research Council of Thailand. Plant Breeding and Multiplication Association of Thailand. Appendices 87
Dr. Phonphimon Wongthida Animal Biotechnology Research Unit The 2014 NRCT PhD Dissertation Award (Medical Science Field) for “Understanding, and Exploiting, the Contribution of the Immune System to the Therapeutic Efficacy of Oncolytic Virotherapy with Vesicular Stomatitis Virus”, awarded by the National Research Council of Thailand.
Dr. Peera Jaruampornpan Animal Biotechnology Research Unit The 2014 NRCT PhD Dissertation Award (Chemistry and Pharmacy Field) for “Post - Translational Membrane Protein Targeting by the Chloroplast Signal Recognition Particle”, awarded by the National Research Council of Thailand.
Dr. Pornkamol Unrean Biochemical Engineering and Pilot Plant Research and Development Laboratory F The 2014 TRF-CHE-Scopus Young Researcher Award (Engineering & Multidisciplinary Technology) for her work on “Systematic Bioprocess Development of Sustainable Biofuel Production”, awarded by the Thailand Research Fund (TRF), Office of the Higher Education Commission (CHE) and Elsevier.
Ms. Phongphan Ek-arwut Rural Development Technology Service Unit An award for her contribution to education in the monastic schools, presented by the National Office of Buddhism. 88 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)
Executives and Management Team Executive Board
Advisors Naksitte Coovattanachai Advisor to Secretary-General, National Science Technology and Innovation Policy Office (STI) Morakot Tanticharoen Professor Emeritus, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi Sermpol Ratasuk Expert on organization planning and environmental impact assessment
Chairman Sakarindr Bhumiratana President, King Mongkut’s University of Technology Thonburi
Vice Chairman Thaweesak Koanantakool President, National Science and Technology Development Agency (NSTDA)
Members Porametee Vimolsiri Deputy Secretary General, Office of the National Economic and Social Development Board (NESDB) Suwit Chaikiattiyos Deputy Director-General, Department of Agriculture Chanun Puttamilinpratheap Deputy Director, Bureau of the Budget Prapon Wilairat Professor Emeritus, Faculty of Science, Mahidol University Amaret Bhumiratana Director, Royal Golden Jubilee Program, Thailand Research Fund (TRF) Julapark Chunwongse Associate Professor, Faculty of Agriculture, Kasetsart University Pornsilp Patcharintanakul Vice Chairman, Thai Chamber of Commerce Deputy Secretary-General, Board of Trade of Thailand Vice President, Charoen Pokphand Group Pachok Pongpanich Executive Committee, Thai Seed Trade Association -THASTA Managing Director, PacThai (Pacific Seeds Co., Ltd.) Rutjawate Taharnklaew Assistant Vice President, R&D Center, Betagro Group Kittiphong Limsuwannarot Vice President of Green Chemicals, PTT Global Chemical Public Company Limited Kanyawim Kirtikara Executive Director, BIOTEC Dussadee Siamhan Deputy Executive Director, BIOTEC Appendices 89
International Advisory Board
Chairman Lene Lange Professor in Chemical and Biochemical Engineering, Technical University of Denmark, DENMARK Members Philippe Desmeth President, World Federation for Culture Collections Martin Keller Associate Laboratory Director of Biological and Environmental Sciences, Oak Ridge National Laboratory (ORNL), USA Vítor Martins dos Santos Chair for Systems and Synthetic Biology, Wageningen University, THE NETHERLANDS Ray Ming Professor of Plant Biology, University of Illinois at Urbana-Champaign, USA Jean-Marcel Ribaut Director, Generation Challenge Programme (GCP) Jonathan V. Sweedler James R. Eiszner Family Chair in Chemistry, University of Illinois Urbana- Champaign, USA Frédéric Tangy Director of Research, French National Centre for Scientific Research (CNRS) and Head of the Viral Genomics and Vaccination Unit, Institut Pasteur, FRANCE Management Team
Kanyawim Kirtikara FExecutive Director
Dussadee Siamhan Wonnop Visessanguan Deputy Executive Director Director, Food Biotechnology Research Unit
Suvit Tia Nitsara Karoonuthaisiri Deputy Executive Director Director, Biosensing Technology Research Unit
Lily Eurwilaichitr Deputy Executive Director and Director, Bioresources Technology Research Unit
Somvong Tragoonrung Director, Genome Technology Research Unit
Sumalee Kamchonwongpaisan Director, Medical Molecular Biology Research Unit
Sirawut Klinbunga Director, Animal Biotechnology Research Unit 90 Annual Report 2015 National Center for Genetic Engineering and Biotechnology (BIOTEC)