DOCSLIB.ORG

Phylogenomic Classification and Biosynthetic Potential of the Fossil

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Phylogenomic Classification and Biosynthetic Potential of the Fossil Phylogenomic Classification and Biosynthetic Potential of the Fossil Fuel-Biodesulfurizing Rhodococcus Strain IGTS8 Dean Thompson, Valérie Cognat, Michael Goodfellow, Sandrine Koechler, Dimitri Heintz, Christine Carapito, Alain van Dorsselaer, Huda Mahmoud, Vartul Sangal, Wael Ismail To cite this version: Dean Thompson, Valérie Cognat, Michael Goodfellow, Sandrine Koechler, Dimitri Heintz, et al.. Phy- logenomic Classification and Biosynthetic Potential of the Fossil Fuel-Biodesulfurizing Rhodococcus Strain IGTS8. Frontiers in Microbiology, Frontiers Media, 2020, 11, 10.3389/fmicb.2020.01417. hal- 03024721 HAL Id: hal-03024721 https://hal.archives-ouvertes.fr/hal-03024721 Submitted on 25 Nov 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. fmicb-11-01417 July 4, 2020 Time: 17:41 # 1 ORIGINAL RESEARCH published: 07 July 2020 doi: 10.3389/fmicb.2020.01417 Phylogenomic Classification and Biosynthetic Potential of the Fossil Fuel-Biodesulfurizing Rhodococcus Strain IGTS8 Dean Thompson1†, Valérie Cognat2†, Michael Goodfellow3, Sandrine Koechler2, Dimitri Heintz2, Christine Carapito4, Alain Van Dorsselaer4, Huda Mahmoud5, Vartul Sangal1 and Wael Ismail6* 1 Faculty of Health and Life Sciences, Northumbria University, Newcastle upon Tyne, United Kingdom, 2 Institut de Biologie Moléculaire des Plantes, Centre National de Recherche Scientifique (CNRS), Université de Strasbourg, Strasbourg, France, 3 School of Natural and Environmental Sciences, Newcastle University, Newcastle upon Tyne, United Kingdom, 4 Laboratoire de Spectrométrie de Masse Bio-organique, Institut Pluridisciplinaire Hubert Curien, UMR 7178 CNRS, Université Edited by: de Strasbourg, Strasbourg, France, 5 Department of Biological Sciences, College of Science, Kuwait University, Safat, Kian Mau Goh, Kuwait, 6 Environmental Biotechnology Program, Life Sciences Department, College of Graduate Studies, Arabian Gulf University of Technology Malaysia, University, Manama, Bahrain Malaysia Reviewed by: Benjamin C. Stark, Rhodococcus strain IGTS8 is the most extensively studied model bacterium for Illinois Institute of Technology, biodesulfurization of fossil fuels via the non–destructive sulfur–specific 4S pathway. This United States Juana Maria Navarro Llorens, strain was initially assigned to Rhodococcus rhodochrous and later to Rhodococcus Complutense University of Madrid, erythropolis thus making its taxonomic status debatable and reflecting the limited Spain resolution of methods available at the time. In this study, phylogenomic analyses of John Joseph Kilbane, Illinois Institute of Technology, the whole genome sequences of strain IGTS8 and closely related rhodococci showed United States that R. erythropolis and Rhodococcus qingshengii are very closely related species, that *Correspondence: Rhodococcus strain IGTS8 is a R. qingshengii strain and that several strains identified Wael Ismail [email protected] as R. erythropolis should be re-classified as R. qingshengii. The genomes of strains †These authors have contributed assigned to these species contain potentially novel biosynthetic gene clusters showing equally to this work that members of these taxa should be given greater importance in the search for new antimicrobials and other industrially important biomolecules. The plasmid-borne dsz Specialty section: This article was submitted to operon encoding fossil fuel desulfurization enzymes was present in R. qingshengii IGTS8 Microbiotechnology, and R. erythropolis XP suggesting that it might be transferable between members of a section of the journal Frontiers in Microbiology these species. Received: 19 April 2020 Keywords: biodesulfurization, Rhodococcus, 4S pathway, phylogenomics, dibenzothiophene, average nucleotide Accepted: 02 June 2020 identity, digital DNA-DNA hybridization Published: 07 July 2020 Citation: Thompson D, Cognat V, INTRODUCTION Goodfellow M, Koechler S, Heintz D, Carapito C, Van Dorsselaer A, The implementation of green and economic technologies in the petroleum industry is of increasing Mahmoud H, Sangal V and Ismail W interest. This is particularly important in view of the continuously increasing global demand (2020) Phylogenomic Classification for cleaner fuels, stricter environmental regulations and dwindling oil prices (Sahu et al., 2015; and Biosynthetic Potential of the Head and Gray, 2016; Ismail et al., 2017). The biotechnological desulfurization of crude oil and Fossil Fuel-Biodesulfurizing transportation fuels, such as diesel and gasoline, holds the potential to overcome or mitigate the Rhodococcus Strain IGTS8. Front. Microbiol. 11:1417. technical, economic and environmental hurdles of conventional thermochemical desulfurization doi: 10.3389/fmicb.2020.01417 technologies (Kilbane, 2006; Mohebali and Ball, 2016). Rhodococcus strain IGTS8 (ATCC 53968) Frontiers in Microbiology| www.frontiersin.org 1 July 2020| Volume 11| Article 1417 fmicb-11-01417 July 4, 2020 Time: 17:41 # 2 Thompson et al. Genome Sequence of Rhodococcus Strain IGTS8 was the first bacterium to be isolated based on its unique ability with those of R. erythropolis and R. qingshengii strains to establish to break C-S bonds of fuel-born organosulfur compounds and its taxonomic status and to compare the biosynthetic and utilize them as a sole sulfur source (Denome et al., 1994). This biodesulfurization capabilities of strains assigned to these species. discovery raised the prospect of using biocatalysts for removal of sulfur from fossil fuels via biodesulfurization (Kilbane and Bielaga, 1990; Kilbane II and Jackowski, 1992). MATERIALS AND METHODS The biodesulfurization reactions are catalyzed by two monooxygenases (DszC and DszA) and a desulfinase (DszB) Bacterial Strain which constitute the non-destructive 4S pathway. This pathway Rhodococcus strain IGTS8 (American Type Culture Collection, was first reported and characterized in the IGTS8 strain which ATCC 53968) was isolated by Kilbane and colleagues at the for many years has served as the most commonly adopted model Gas Technology Institute-Chicago based on its sulfur-specific for biodesulfurization studies. The genes of the 4S pathway, dszA, fuel biodesulfurization capabilities (Kilbane and Bielaga, 1990; dszB, and dszC, are carried as an operon on a linear megaplasmid Kilbane II and Jackowski, 1992). in the IGTS8 strain (Kilbane and Stark, 2016). These genes have been found in R. erythropolis strains A66 and A69 which Genomic DNA Extraction and use various organosulfur and organonitrogen compounds as Sequencing sole sources of sulfur and nitrogen, respectively (Santos et al., Rhodococcus strain IGTS8 was grown overnight in nutrient 2007). DszC shows sulfur acquisition oxidoreductase (SfnB ◦ family) and acyl-CoA dehydrogenase activities and catalyzes broth at 28 C and the cells pelleted by centrifugation. Genomic the conversion of dibenzothiophene to dibenzothiophene DNA was extracted with the Wizard Genomic DNA purification sulfone (Gray et al., 1996). DszA, a FMN-dependent kit (Promega) using the manufacturer’s instructions for Gram oxidoreductase of the nitrilotriacetate monooxygenase family, positive bacteria and a library constructed with the Nextera converts dibenzothiophene sulfone to 2-(20-hydroxyphenyl) XT DNA Library Prep kit (Ilumina) using 1 ng of genomic benzenesulfinate which is catalytically converted to DNA. The library was evaluated with a 2100 Bioanalyzer (Agilent 2-hydroxybiphenyl and sulfite by the desulfinase DszB. Technologies) before sequencing on an Illumina Miseq system × Rhodococcus strain IGTS8 was classified as Rhodococcus (2 150 paired end reads). rhodochrous (Zopf, 1891; Tsukamura, 1974) by Kayser et al. (1993) prior to being reclassified as R. erythropolis (Gray and Genome Assembly and Computational Thornton, 1928; Goodfellow and Alderson, 1977; Oldfield Analyses et al., 1998; Santos et al., 2007). These initial classifications The PCR duplicates in the libraries were filtered with FastUniq likely reflect the limited resolution of the taxonomic methods 1.1 (Xu et al., 2012) and low-quality (<30) and adapter sequences available at the time (Goodfellow et al., 1998, 1999). Members trimmed with cutadapt 1.14 (Martin, 2011). Contaminant of the genus Rhodococcus (Zopf, 1891) emend Goodfellow screening was performed with FastQ Screen 0.11.1 (Wingett and et al.(1998) are known for their metabolic versatility and Andrews, 2018) to filter reads mapping on the PhiX sequence, potential biotechnological applications in areas such as vector sequences in the NCBI UniVec database1 and other species bioremediation, biotransformation and biocatalysis. They including Homo sapiens (hg19), Arabidopsis thaliana (TAIR10), also produce biosurfactants, carotenoids and bioflocculation Escherichia coli (NC_000913.3), Stenotrophomonas maltophilia agents of industrial interest (Gürtler and Seviour, 2010; Jones (NC_010943.1) and Pseudomonas aeruginosa (NC_002516.2). and Goodfellow, 2012). The remaining singleton reads in the data were excluded with The extent of the heterogeneity encompassed within the genus the tool makepairs of the Pairfq
Load more

Recommended publications
  • Polyphasic Classification of the Gifted Natural Product Producer Streptomyces
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Digital.CSIC © Van der Aart, L.T., Nouioui, I., Kloosterman, A., Igual, J.M., Willemse, J., Goodfellow, M., van Wezel, J.P. 2019. The definitive peer reviewed, edited version of this article is published in International Journal of Systematic and Evolutionary Microbiology, 69, 4, 2019, http://dx.doi.org/10.1099/ijsem.0.003215 Polyphasic classification of the gifted natural product producer Streptomyces roseifaciens sp. nov. Lizah T. van der Aart 1, Imen Nouioui 2, Alexander Kloosterman 1, José Mariano Ingual 3, Joost Willemse 1, Michael Goodfellow 2, *, Gilles P. van Wezel 1,4 *. 1 Molecular Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands 2 School of Natural and Environmental Sciences, University of Newcastle, Newcastle upon Tyne NE1 7RU, UK. 3 Instituto de Recursos Naturales y Agrobiologia de Salamanca, Consejo Superior de Investigaciones Cientificas (IRNASACSIC), c/Cordel de Merinas 40-52, 37008 Salamanca, Spain 4: Department of Microbial Ecology, Netherlands, Institute of Ecology (NIOO-KNAW) Droevendaalsteeg 10, Wageningen 6708 PB, The Netherlands *Corresponding authors. Michael Goodfellow: [email protected], Tel: +44 191 2087706. Gilles van Wezel: Email: [email protected], Tel: +31 715274310. Accession for the full genome assembly: GCF_001445655.1 1 Abstract A polyphasic study was designed to establish the taxonomic status of a Streptomyces strain isolated from soil from the QinLing Mountains, Shaanxi Province, China, and found to be the source of known and new specialized metabolites. Strain MBT76 T was found to have chemotaxonomic, cultural and morphological properties consistent with its classification in the genus Streptomyces .
    [Show full text]
  • INVESTIGATING the ACTINOMYCETE DIVERSITY INSIDE the HINDGUT of an INDIGENOUS TERMITE, Microhodotermes Viator
    INVESTIGATING THE ACTINOMYCETE DIVERSITY INSIDE THE HINDGUT OF AN INDIGENOUS TERMITE, Microhodotermes viator by Jeffrey Rohland Thesis presented for the degree of Doctor of Philosophy in the Department of Molecular and Cell Biology, Faculty of Science, University of Cape Town, South Africa. April 2010 ACKNOWLEDGEMENTS Firstly and most importantly, I would like to thank my supervisor, Dr Paul Meyers. I have been in his lab since my Honours year, and he has always been a constant source of guidance, help and encouragement during all my years at UCT. His serious discussion of project related matters and also his lighter side and sense of humour have made the work that I have done a growing and learning experience, but also one that has been really enjoyable. I look up to him as a role model and mentor and acknowledge his contribution to making me the best possible researcher that I can be. Thank-you to all the members of Lab 202, past and present (especially to Gareth Everest – who was with me from the start), for all their help and advice and for making the lab a home away from home and generally a great place to work. I would also like to thank Di James and Bruna Galvão for all their help with the vast quantities of sequencing done during this project, and Dr Bronwyn Kirby for her help with the statistical analyses. Also, I must acknowledge Miranda Waldron and Mohammed Jaffer of the Electron Microsope Unit at the University of Cape Town for their help with scanning electron microscopy and transmission electron microscopy related matters, respectively.
    [Show full text]
  • The Genome Analysis of the Human Lung-Associated Streptomyces Sp
    microorganisms Article The Genome Analysis of the Human Lung-Associated Streptomyces sp. TR1341 Revealed the Presence of Beneficial Genes for Opportunistic Colonization of Human Tissues Ana Catalina Lara 1,† , Erika Corretto 1,†,‡ , Lucie Kotrbová 1, František Lorenc 1 , KateˇrinaPetˇríˇcková 2,3 , Roman Grabic 4 and Alica Chro ˇnáková 1,* 1 Institute of Soil Biology, Biology Centre Academy of Sciences of The Czech Republic, Na Sádkách 702/7, 37005 Ceskˇ é Budˇejovice,Czech Republic; [email protected] (A.C.L.); [email protected] (E.C.); [email protected] (L.K.); [email protected] (F.L.) 2 Institute of Immunology and Microbiology, 1st Faculty of Medicine, Charles University, Studniˇckova7, 12800 Prague 2, Czech Republic; [email protected] 3 Faculty of Science, University of South Bohemia, Branišovská 1645/31a, 37005 Ceskˇ é Budˇejovice, Czech Republic 4 Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zátiší 728/II, 38925 Vodˇnany, Czech Republic; [email protected] * Correspondence: [email protected] † Both authors contributed equally. ‡ Current address: Faculty of Science and Technology, Free University of Bozen-Bolzano, Universitätsplatz 5—piazza Università 5, 39100 Bozen-Bolzano, Italy. Citation: Lara, A.C.; Corretto, E.; Abstract: Streptomyces sp. TR1341 was isolated from the sputum of a man with a history of lung and Kotrbová, L.; Lorenc, F.; Petˇríˇcková, kidney tuberculosis, recurrent respiratory infections, and COPD. It produces secondary metabolites K.; Grabic, R.; Chroˇnáková,A. associated with cytotoxicity and immune response modulation. In this study, we complement The Genome Analysis of the Human our previous results by identifying the genetic features associated with the production of these Lung-Associated Streptomyces sp.
    [Show full text]
  • Streptomyces Oryzae Sp. Nov., an Endophytic Actinomycete Isolated from Stems of Rice Plant
    The Journal of Antibiotics (2015) 68, 368–372 & 2015 Japan Antibiotics Research Association All rights reserved 0021-8820/15 www.nature.com/ja ORIGINAL ARTICLE Streptomyces oryzae sp. nov., an endophytic actinomycete isolated from stems of rice plant Ratchanee Mingma1,2, Kannika Duangmal1,2, Arinthip Thamchaipenet2,3, Savitr Trakulnaleamsai1,2, Atsuko Matsumoto4 and Yoko Takahashi4 An actinomycete strain S16–07T, isolated from surface-sterilized stems of rice plant (Oryza sativa L.), was characterized using a polyphasic approach. Phylogenetic analysis of 16S rRNA gene sequences indicated affiliation of the strain belonged to the genus Streptomyces. The highest levels of sequence similarity were found with Streptomyces smyrnaeus SM3501T (97.7% similarity), S. abikoensis NBRC 13860T (97.6% similarity) and S. thermocarboxydovorans NBRC 16324T (97.5% similarity). The cell wall T of strain S16–07 contained LL-diaminopimelic acid. The predominant menaquinones were MK-9(H6) and MK-9(H8). Phospholipids detected were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, hydroxy- phosphatidylethanolamine, hydroxy-phosphatidylmonomethylethanolamine and phosphatidylinositol mannosides. The major T cellular fatty acids were ai-C15:0, i-C16:0 and ai-C17:0. The G+C content of strain S16–07 was 70.4 mol%. On the basis of the phylogeny of the isolate and its differences from the most closely related species, the isolate S16–07T represents a novel species for which the name S. oryzae sp. nov. is proposed. The type strain is S16–07T ( = BCC 60400T = NBRC 109761T). The Journal of Antibiotics (2015) 68, 368–372; doi:10.1038/ja.2014.166; published online 14 January 2015 INTRODUCTION MATERIALS AND METHODS The genus Streptomyces was first proposed by Waksman and Strain S16–07T was isolated from surface-sterilized stems of rice plant Henrici1 and was classified in the family Streptomycetaceae,order (O.
    [Show full text]
  • Antifungal Activity and Biosynthetic Potential of New Streptomyces Sp
    International Journal of Molecular Sciences Article Antifungal Activity and Biosynthetic Potential of New Streptomyces sp. MW-W600-10 Strain Isolated from Coal Mine Water Piotr Siupka 1,* , Frederik Teilfeldt Hansen 2, Aleksandra Schier 1, Simone Rocco 1, Trine Sørensen 2 and Zofia Piotrowska-Seget 1 1 Faculty of Natural Sciences, Institute of Biology, Biotechnology and Environmental Protection, University of Silesia in Katowice, 40032 Katowice, Poland; [email protected] (A.S.); [email protected] (S.R.); zofi[email protected] (Z.P.-S.) 2 Faculty of Engineering and Science, Department of Chemistry and Biosciences, University of Aalborg, 9220 Aalborg, Denmark; [email protected] (F.T.H.); [email protected] (T.S.) * Correspondence: [email protected] Abstract: Crop infections by fungi lead to severe losses in food production and pose risks for human health. The increasing resistance of pathogens to fungicides has led to the higher usage of these chemicals, which burdens the environment and highlights the need to find novel natural biocontrol agents. Members of the genus Streptomyces are known to produce a plethora of bioactive compounds. Recently, researchers have turned to extreme and previously unexplored niches in the search for new strains with antimicrobial activities. One such niche are underground coal mine environments. We isolated the new Streptomyces sp. MW-W600-10 strain from coal mine water samples collected at 665 m below ground level. We examined the antifungal activity of the strain against plant pathogens Citation: Siupka, P.; Hansen, F.T.; Fusarium culmorum DSM62188 and Nigrospora oryzae roseF7. Furthermore, we analyzed the strain’s Schier, A.; Rocco, S.; Sørensen, T.; biosynthetic potential with the antiSMASH tool.
    [Show full text]
  • Marine Actinobacteria As a Source of Compounds for Phytopathogen Control: an Integrative Metabolic-Profiling / Bioactivity and Taxonomical Approach
    RESEARCH ARTICLE Marine Actinobacteria as a source of compounds for phytopathogen control: An integrative metabolic-profiling / bioactivity and taxonomical approach Luz A. Betancur1,2, Sandra J. Naranjo-Gaybor1,3, Diana M. Vinchira-Villarraga1, Nubia C. Moreno-Sarmiento1, Luis A. Maldonado4, Zulma R. Suarez-Moreno5, Alejandro Acosta- GonzaÂlez6, Gillermo F. Padilla-Gonzalez7, Mo nica Puyana8, Leonardo Castellanos1, a1111111111 Freddy A. Ramos1* a1111111111 a1111111111 1 Universidad Nacional de Colombia, Sede BogotaÂ, Departamento de QuõÂmica, Carrera, Edificio de QuõÂmica of 427, BogotaÂ, Colombia, 2 Universidad de Caldas. Departamento de QuõÂmica. Edificio Orlando Sierra, a1111111111 Bloque B, Sede Palogrande Calle. Manizales, Caldas, Colombia, 3 Universidad de las Fuerzas Armadas, a1111111111 ESPE Carrera de IngenierõÂa Agropecuaria IASA II Av. General Rumiñahui s/n, SangolquõÂ- Ecuador, 4 Universidad AutoÂnoma Metropolitana RectorõÂaÐSecretarõÂa General, ProlongacioÂn Canal de Miramontes, Col. Ex-hacienda San Juan de Dios, Tlalpan, MeÂxico DF, 5 InvestigacioÂn y Desarrollo, Empresa Colombiana de Productos Veterinarios VECOL S.A., Bogota D.C, 6 Universidad de la Sabana, Facultad de IngenierõÂa, Autopista Norte, ChõÂa, Cundinamarca, Colombia, 7 Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Av. do de Sao Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão OPEN ACCESS Preto, Av. do CafeÂ, Ribeirão Preto±SP, Brazil, 8 Departamento de Ciencias BioloÂgicas y Ambientales, Citation: Betancur LA, Naranjo-Gaybor SJ, Programa de BiologõÂa Marina, Universidad Jorge Tadeo Lozano, Carrera, Modulo, Oficina, BogotaÂ, Colombia Vinchira-Villarraga DM, Moreno-Sarmiento NC, * [email protected] Maldonado LA, Suarez-Moreno ZR, et al. (2017) Marine Actinobacteria as a source of compounds for phytopathogen control: An integrative metabolic-profiling / bioactivity and taxonomical Abstract approach.
    [Show full text]
  • Dissimilatory Nitrate Reduction in Bacillus Copyright ©2017, Yihua Sun ISBN-Number: 978-94-6197-486-0 All Rights Are Reserved
    Dissimilatory nitrate reduction in Bacillus Yihua Sun Promotors Dr. Kim Heylen Prof. Dr. em. Paul De Vos Prof. Dr. Anne Willems Dissertation submitted in fulfilment of the requirements for the degree of Doctor (Ph.D.) of Science: Biotechnology (Ghent University) Yihua Sun – Dissimilatory nitrate reduction in Bacillus Copyright ©2017, Yihua Sun ISBN-number: 978-94-6197-486-0 All rights are reserved. No part of this thesis protected by this copyright notice may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording or by any information storage or retrieval system without written permission of the author and promotors. Printed by University Press | www.universitypress.be Ph.D. thesis, Faculty of Sciences, Ghent University, Ghent, Belgium. This Ph.D. work was financially supported by Chinese scholar council ( 201206330054) and BOF CSC co- funding from Ghent University (01SC2713) Publicly defended in Ghent, Belgium, January 20 th , 2017 Examination committee Prof. Dr. Savvas Savvides (Chairman) L-Probe: Laboratory for Protein Biochemistry and Biomolecular Engineering Faculty of Sciences, Ghent University, Belgium VIB Inflammation Research Center VIB, Ghent, Belgium Prof. Dr. Anne Willems (Promotor) LM-UGent: Laboratory of Microbiology Faculty of Sciences, Ghent University, Belgium Prof. Dr. em. Paul De Vos (Promotor) LM-UGent: Laboratory of Microbiology Faculty of Sciences, Ghent University, Belgium Dr. Kim Heylen (Promotor) LM-UGent: Laboratory of Microbiology Faculty of Sciences, Ghent University, Belgium Prof. Dr. Sofie Goormachtig (Secretary) PSB: Plant systems Biology Faculty of Sciences, VIB, Ghent University, Belgium Prof. Dr. ir. Nico Boon CMET: Center for Microbial Ecology and Technology Faculty of Bioscience Engineering, Ghent University, Belgium Prof.
    [Show full text]
  • Nitration Enzyme Toolkit for the Biosynthesis of Energetic Materials
    ORNL/SPR-2017/498 Oak Ridge National Laboratory Nitration enzyme toolkit for the biosynthesis of energetic materials PI: David E. Graham SERDP Project Number Jim C. Spain WP-2332 Ronald J. Parry Robert L. Hettich February 5, 2018 Approved for public release. Distribution is unlimited. Form Approved REPORT DOCUMENTATION PAGE OMB No. 0704-0188 Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services, Directorate for Information Operations and Reports (0704-0188), 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202- 4302. Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE (DD-MM-YYYY) 2. REPORT TYPE 3. DATES COVERED (From - To) 02-05-2018 Final Report 01-09-2013 to 09-30-2017 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER WP-2332 Nitration enzyme toolkit for the biosynthesis of energetic materials 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) 5d. PROJECT NUMBER Graham, David E; Spain, Jim C; Parry, Ronald J; Hettich, Robert L; Mahan, Kristina M; 5e.
    [Show full text]
  • A Novel Diterpene Agent Isolated from Microbispora Hainanensis Strain
    www.nature.com/scientificreports OPEN A novel diterpene agent isolated from Microbispora hainanensis strain CSR‑4 and its in vitro and in silico inhibition efects on acetylcholine esterase enzyme Chitti Thawai1,2,3*, Nantiya Bunbamrung4, Pattama Pittayakhajonwut4, Sumet Chongruchiroj5, Jaturong Pratuangdejkul5, Ya‑Wen He6, Sarin Tadtong7, Vipaporn Sareedenchai7, Pinidphon Prombutara8 & Yang Qian9 An actinomycete strain CSR‑4 was isolated from the rhizosphere soil of Zingiber montanum. Taxonomic characterization revealed strain CSR‑4 was a member of the genus Microbispora. Whole‑ genome sequence analysis exhibited the highest average nucleotide identity (ANI) value (95.34%) and digital DNA–DNA hybridization (DDH) value (74.7%) between strain CSR‑4 and the closest relative M. hainanensis DSM 45428T, which was in line with the assignment to same species. In addition, a new diterpene compound, 2α‑hydroxy‑8(14), 15‑pimaradien‑17, 18‑dioic acid, and nine known compounds were isolated from the ethyl acetate crude extract of fermentation broth. Interestingly, a new diterpene displayed the suppressive efect on the recombinant human acetylcholinesterase (rhAChE) enzymes (IC50 96.87 ± 2.31 μg/ml). In silico studies based on molecular docking and molecular dynamics (MD) simulations were performed to predict a binding mode of the new compound into the binding pocket of the rhAChE enzyme and revealed that some amino acids in the peripheral anions site (PAS), anionic subsite, oxyanion site and catalytic active site (CAS) of the rhAChE have interacted with the compound. Therefore, our new compound could be proposed as a potential active human AChE inhibitor. Moreover, the new compound can protect signifcantly the neuron cells (% neuron viability = 88.56 ± 5.19%) from oxidative stress induced by serum deprivation method at 1 ng/ml without both neurotoxicities on murine P19‑derived neuron cells and cytotoxicity against Vero cells.
    [Show full text]
  • Antimicrobial Activity of Streptomyces Spp. Isolated from Apis Dorsata Combs Against Some Phytopathogenic Bacteria
    Antimicrobial activity of Streptomyces spp. isolated from Apis dorsata combs against some phytopathogenic bacteria Yaowanoot Promnuan1, Saran Promsai1 and Sujinan Meelai2 1 Department of Microbiology, Faculty of Liberal Arts and Science, Kasetsart University-Kamphaeng Saen campus, Kamphaeng Saen, Nakhon Pathom, Thailand 2 Department of Microbiology, Faculty of Science, Silpakorn University-Sanam Chandra Palace campus, Nakhon Pathom, Nakhon Pathom, Thailand ABSTRACT The aim of this study was to investigate the antimicrobial potential of actinomycetes isolated from combs of the giant honey bee, Apis dorsata. In total, 25 isolates were obtained from three different media and were screened for antimicrobial activity against four plant pathogenic bacteria (Ralstonia solanacearum, Xanthomonas campestris pv. campestris, Xanthomonas oryzae pv. oryzae and Pectobacterium carotovorum). Following screening using a cross-streaking method, three isolates showed the potential to inhibit the growth of plant pathogenic bacteria. Based on a 96-well microtiter assay, the crude extract of DSC3-6 had minimum inhibitory concentration (MIC) values against X. oryzae pv. oryzae, X. campestris pv. campestris, R. solanacearum and P. carotovorum of 16, 32, 32 and 64 mg L−1, respectively. The crude extract of DGA3-20 had MIC values against X. oryzae pv. oryzae, X. campestris pv. campestris, R. solanacearum and P. carotovorum of 32, 32, 32 and 64 mg L−1, respectively. The crude extract of DGA8-3 at 32 mgL−1 inhibited the growth of X. oryzae pv. oryzae, X. campestris pv. campestris, R. solanacearum and P. carotovorum. Based on their 16S rRNA gene sequences, all isolates were identified as members of the genus Streptomyces. The analysis of 16S rRNA gene sequence similarity and of the phylogenetic tree based on the maximum likelihood algorithm showed that isolates DSC3-6, DGA3-20 and DGA8-3 were closely related Submitted 30 April 2020 to Streptomyces ramulosus (99.42%), Streptomyces axinellae (99.70%) and Streptomyces Accepted 16 November 2020 drozdowiczii (99.71%), respectively.
    [Show full text]