DOCSLIB.ORG

Metabolic Engineering and Systems Biology for Increasing Biofuel Production in Microalgae

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Metabolic Engineering and Systems Biology for Increasing Biofuel Production in Microalgae METABOLIC ENGINEERING AND SYSTEMS BIOLOGY FOR INCREASING BIOFUEL PRODUCTION IN MICROALGAE by Robert Edward Jinkerson Copyright by Robert E. Jinkerson 2013 Some Rights Reserved This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License http://creativecommons.org/licenses/by-nc-sa/4.0/ A thesis submitted to the Faculty and the Board of Trustees of the Colorado School of Mines in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Applied Chemistry). Golden, Colorado Date ___________ Signed: _______________________ Robert Jinkerson Signed: _______________________ Dr. Matthew C. Posewitz Thesis Advisor Golden, Colorado Date ___________ Signed: _______________________ Dr. David T. Wu Professer and Head Department of Chemistry and Geochemistry ii ABSTRACT The saccharification of starch coupled with fermentation to ethanol and the transesterification of vegetable oils to produce biodiesel are mature technologies that currently provide the majority of biofuels in the United States. However, traditional food-based starch and oil feedstocks, such as corn and soybean, cannot meet our current fuel demands and their use remains controversial. Microalgae have been of recent interest for use as a biofuel feedstock because they can produce large quantities of carbohydrates and lipids, contain little recalcitrant biomass, and do not impact the food supply. In the green alga Chlamydomonas reinhardtii, the primary carbohydrate produced is starch and the primary storage lipid produced is triacylglyceride (TAG), but high yields of these bioenergy carriers are usually only found under conditions of nutrient stress (N, S, P). As a strategy for increasing TAG yields, starchless mutants that divert carbon away from starch biosynthesis and into TAG production were investigated. These starchless mutants produce more TAG than wildtype, but at the expense of total productivity and lower overall anabolic activity. To increase starch levels, a native enzyme key to starch biosynthesis, isoamylase 1 (ISA1), was introduced into these algae which resulted in starch excess phenotypes. These mutant strains accumulate 3 to 4 fold more total glucan under nutrient-replete conditions by diverting metabolic flux into starch biosynthesis at the expense of cell division and protein synthesis. The starches produced by these algae are more crystalline and larger in size than wildtype. None of the current genetically-tractable model algal species are competitive TAG production strains. To alleviate this the commercially cultivated, oleaginous alga Nannochloropsis gaditana CCMP526 was developed into a new model algal species for investigating TAG production. The genome and transcriptome was sequence and assembled, gene models developed, and metabolic pathways in this organism were reconstructed. Phylogenomic analysis identified genetic attributes of this organism, including gene expansions and unique stramenopile photosynthesis genes, that may explain the distinguishing oleaginous phenotypes observed. The availability of a genome sequence and transformation method are facilitating investigations into N. gaditana lipid biosynthesis and will permit genetic engineering strategies to further improve this naturally productive algal strain. iii TABLE OF CONTENTS ABSTRACT ................................................................................................................................................ iii LIST OF FIGURES ..................................................................................................................................... ix LIST OF TABLES ....................................................................................................................................... xi LIST OF ABBREVIATIONS ..................................................................................................................... xii ACKNOWLEDGMENTS .......................................................................................................................... xv DEDICATION ........................................................................................................................................... xvi CHAPTER 1 Introduction ............................................................................................................................. 1 1.1 Biofuels overview...................................................................................................................... 1 1.2 Microalgae as a biofuel feedstock ............................................................................................. 4 1.3 Algal carbohydrates ................................................................................................................... 7 1.3.1 Polyglucan storage and starch biosynthesis in algae ......................................................... 8 1.3.2 Starch degradation in algae ............................................................................................... 9 1.3.3 Genetic modification strategies for increasing glucan storage ........................................ 11 1.3.4 Genetic modification strategies for decreasing starch degradation in microalgae .......... 11 1.3.5 Secretion strategies and soluble sugars ........................................................................... 12 1.3.6 Alcohol production in algae ............................................................................................ 13 1.4 Algal lipids .............................................................................................................................. 13 1.4.1 Fatty acids ....................................................................................................................... 14 1.4.2 Altering algal fatty acid biosynthesis for biofuel production .......................................... 14 1.4.3 Terpenes and terpenoids .................................................................................................. 16 1.4.4 Botryococcus hydrocarbons ............................................................................................ 16 1.5 The focus of this work ............................................................................................................. 18 1.5.1 Carbon partitioning in green algae between starch and lipids ........................................ 18 1.5.2 Role of isoamylase in starch biosynthesis and in starch excess phenotype .................... 19 1.5.3 How systems biology can used to increase biofuel production in algal systems ............ 20 1.5.4 Development of a new model organism for algal lipid production, Nannochloropsis gaditana ................................................................................................................................... 21 1.6 General summary .................................................................................................................... 23 CHAPTER 2 Increased Lipid Accumulation in the Chlamydomonas reinhardtii sta7-10 Starchless Isoamylase Mutant and Increased Carbohydrate Synthesis in Complemented Strains ......... 24 2.1 Introduction ............................................................................................................................. 24 2.2 Material and methods .............................................................................................................. 25 2.2.1 Chlorophyll measurements. ............................................................................................ 26 iv 2.2.2 Microscopy ..................................................................................................................... 26 2.2.3 Starch assays ................................................................................................................... 27 2.2.4 FAME Quantification ..................................................................................................... 27 2.2.5 Acetate utilization ........................................................................................................... 28 2.2.6 Photosynthetic oxygen evolution rates ............................................................................ 28 2.2.7 Chlorophyll Fluorescence Measurements ....................................................................... 28 2.3 Results ..................................................................................................................................... 29 2.4 Discussion ............................................................................................................................... 39 2.5 Conclusion ............................................................................................................................... 43 CHAPTER 3 Isoamylase 1: single gene starch trigger in green algae ........................................................ 44 3.1 Introduction ............................................................................................................................. 44 3.2 Methods and Materials ............................................................................................................ 44 3.2.1 Strains, culturing conditions, growth assessments .......................................................... 45 3.2.2 ISA1 Transformation ....................................................................................................... 45 3.2.3 Starch quantification ......................................................................................................
Load more

Recommended publications
  • Open 1-Dissertation NEK V12 4
    The Pennsylvania State University The Graduate School Department of Chemical Engineering DEVELOPMENT OF BIOLOGICAL PLATFORM FOR THE AUTOTROPHIC PRODUCTION OF BIOFUELS A Dissertation in Chemical Engineering by Nymul Khan 2015 Nymul Khan Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy May 2015 The dissertation of Nymul Khan was reviewed and approved* by the following: Wayne Curtis Professor, Chemical Engineering Dissertation Advisor Chair of Committee Esther Gomez Assistant Professor, Chemical Engineering Manish Kumar Assistant Professor, Chemical Engineering John Regan Professor, Environmental Engineering Phillip E. Savage Walter L. Robb Family Endowed Chair Head of the Department of Chemical Engineering *Signatures are on file in the Graduate School iii ABSTRACT The research described herein is aimed at developing an advanced biofuel platform that has the potential to surpass the natural rate of solar energy capture and CO2 fixation. The underlying concept is to use the electricity from a renewable source, such as wind or solar, to capture CO2 via a biological agent, such as a microbe, into liquid fuels that can be used for the transportation sector. In addition to being renewable, the higher rate of energy capture by photovoltaic cells than natural photosynthesis is expected to facilitate higher rate of liquid fuel production than traditional biofuel processes. The envisioned platform is part of ARPA-E’s (Advanced Research Projects Agency - Energy) Electrofuels initiative which aims at supplementing the country’s petroleum based fuel production with renewable liquid fuels that can integrate easily with the existing refining and distribution infrastructure (http://arpa- e.energy.gov/ProgramsProjects/Electrofuels.aspx).
    [Show full text]
  • Properties of Chlorophyllase from Capsicum Annuum L. Fruits
    Properties of Chlorophyllase from Capsicum annuum L. Fruits Dámaso Hornero-Méndez and Marí a Isabel Mínguez-Mosquera* Departamento de Biotecnologia de Alimentos, Instituto de la Grasa (CSIC), Av. Padre Garcia Tejero, 4, 41012-Sevilla, SPAIN. Fax: +34-954691262. E-mail: [email protected] * Author for correspondence and reprint requests Z. Naturforsch. 56c, 1015-1021 (2001); received June 27/August 6 , 2001 Chlorophyll, Chlorophyllase, Capsicum annuum The in vitro properties of semi-purified chlorophyllase (chlorophyll-chlorophyllido hy­ drolase, EC 3.1.1.14) from Capsicum annuum fruits have been studied. The enzyme showed an optimum of activity at pH 8.5 and 50 °C. Substrate specificity was studied for chlorophyll (Chi) a, Chi b, pheophytin (Phe) a and Phe b, with K m values of 10.70, 4.04, 2.67 and 6.37 ^im respectively. Substrate inhibition was found for Phe b at concentrations higher than 5 ^m. Chlorophyllase action on Chi a ’ and Chi b' was also studied but no hydrolysis was observed, suggesting that the mechanism of action depends on the configuration at C-132 in the chloro­ phyll molecule, with the enzyme acting only on compounds with R132 stereochemistry. The effect of various metals (Mg2+, Hg2+, Cu2+, Zn2+, Co , Fe2+ and Fe3+) was also investigated, and a general inhibitory effect was found, this being more marked for Hg2+ and Fe2+. Func­ tional groups such as -SH and -S-S- seemed to participate in the formation of the enzyme- substrate complex. Chelating ion and the carbonyl group at C3 appeared to be important in substrate recognition by the enzyme.
    [Show full text]
  • Photosynthetic Pigments in Diatoms
    Mar. Drugs 2015, 13, 5847-5881; doi:10.3390/md13095847 OPEN ACCESS marine drugs ISSN 1660-3397 www.mdpi.com/journal/marinedrugs Review Photosynthetic Pigments in Diatoms Paulina Kuczynska 1, Malgorzata Jemiola-Rzeminska 1,2 and Kazimierz Strzalka 1,2,* 1 Faculty of Biochemistry, Biophysics and Biotechnology, Department of Plant Physiology and Biochemistry, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland; E-Mails: [email protected] (P.K.); [email protected] (M.J.-R.) 2 Małopolska Centre of Biotechnology, Gronostajowa 7A, Krakow 30-387, Poland * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +48-126-646-509; Fax: +48-126-646-902. Academic Editor: Véronique Martin-Jézéquel Received: 10 July 2015 / Accepted: 7 September 2015 / Published: 16 September 2015 Abstract: Photosynthetic pigments are bioactive compounds of great importance for the food, cosmetic, and pharmaceutical industries. They are not only responsible for capturing solar energy to carry out photosynthesis, but also play a role in photoprotective processes and display antioxidant activity, all of which contribute to effective biomass and oxygen production. Diatoms are organisms of a distinct pigment composition, substantially different from that present in plants. Apart from light-harvesting pigments such as chlorophyll a, chlorophyll c, and fucoxanthin, there is a group of photoprotective carotenoids which includes β-carotene and the xanthophylls, diatoxanthin, diadinoxanthin, violaxanthin, antheraxanthin, and zeaxanthin, which are engaged in the xanthophyll cycle. Additionally, some intermediate products of biosynthetic pathways have been identified in diatoms as well as unusual pigments, e.g., marennine. Marine algae have become widely recognized as a source of unique bioactive compounds for potential industrial, pharmaceutical, and medical applications.
    [Show full text]
  • Structure of Pigment Metabolic Pathways and Their Contributions to White Tepal Color Formation of Chinese Narcissus Tazetta Var
    International Journal of Molecular Sciences Article Structure of Pigment Metabolic Pathways and Their Contributions to White Tepal Color Formation of Chinese Narcissus tazetta var. chinensis cv Jinzhanyintai Yujun Ren † ID , Jingwen Yang †, Bingguo Lu, Yaping Jiang, Haiyang Chen, Yuwei Hong, Binghua Wu and Ying Miao * Center for Molecular Cell and Systems Biology, Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China; [email protected] (Y.R.); [email protected] (J.Y.); [email protected] (B.L.); [email protected] (Y.J.); [email protected] (H.C.); [email protected] (Y.H.); [email protected] (B.W.) * Correspondence: [email protected]; Tel.:/Fax: +86-591-8639-2987 † These authors contributed equally to this work. Received: 7 August 2017; Accepted: 4 September 2017; Published: 8 September 2017 Abstract: Chinese narcissus (Narcissus tazetta var. chinensis) is one of the ten traditional flowers in China and a famous bulb flower in the world flower market. However, only white color tepals are formed in mature flowers of the cultivated varieties, which constrains their applicable occasions. Unfortunately, for lack of genome information of narcissus species, the explanation of tepal color formation of Chinese narcissus is still not clear. Concerning no genome information, the application of transcriptome profile to dissect biological phenomena in plants was reported to be effective. As known, pigments are metabolites of related metabolic pathways, which dominantly decide flower color. In this study, transcriptome profile and pigment metabolite analysis methods were used in the most widely cultivated Chinese narcissus “Jinzhanyintai” to discover the structure of pigment metabolic pathways and their contributions to white tepal color formation during flower development and pigmentation processes.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 9,725,399 B2 Petrie Et Al
    USO09725399B2 (12) United States Patent (10) Patent No.: US 9,725,399 B2 Petrie et al. (45) Date of Patent: Aug. 8, 2017 (54) LPID COMPRISING LONG CHAN (51) Int. Cl. POLYUNSATURATED FATTY ACDS C07C 69/587 (2006.01) CIIB I/O (2006.01) (71) Applicants: Commonwealth Scientific and (Continued) Industrial Research Organisation, (52) U.S. Cl. Acton, Australian Capital Territory CPC .............. C07C 69/587 (2013.01); A23D 9/00 (AU): Nuseed Pty Ltd, Laverton North, (2013.01); A61K 36/31 (2013.01): CIIB I/10 Victoria (AU); Grains Research and (2013.01); A61 K 2.236/00 (2013.01) Development Corporation, Barton, (58) Field of Classification Search Australian Capital Territory (AU) CPC .......................... C12N 15/8247; CO7C 69/587 See application file for complete search history. (72) Inventors: James Robertson Petrie, Goulburn (AU); Surinder Pal Singh, Downer (56) References Cited (AU); Pushkar Shrestha, Lawson U.S. PATENT DOCUMENTS (AU); Jason Timothy McAllister, Portarlington (AU); Robert Charles De 4,399.216 A 8, 1983 Axel et al. Feyter, Monash (AU); Malcolm David 5,004,863. A 4, 1991 Umbeck Devine, Vernon (CA) (Continued) (73) Assignees: COMMONWEALTH SCIENTIFIC FOREIGN PATENT DOCUMENTS AND INDUSTRIAL RESEARCH AU 667939 1, 1994 ORGANISATION, Campbell (AU): AU 200059710 B2 12/2000 NUSEED PTY LTD, Laverton North (Continued) (AU); GRAINS RESEARCH AND DEVELOPMENT CORPORATION, Barton (AU) OTHER PUBLICATIONS Ruiz-Lopez, N. et al., “Metabolic engineering of the omega-3 long (*) Notice: Subject to any disclaimer, the term of this chain polyunsaturated fatty acid biosynthetic pathway into trans patent is extended or adjusted under 35 genic plants' Journal of Experimental botany, 2012, vol.
    [Show full text]
  • Infection by a Giant Virus (Aav) Induces Widespread Physiological Reprogramming in Aureococcus Anophagefferens CCMP1984 – a Harmful Bloom Algae
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Microbiology Publications and Other Works Microbiology 4-19-2018 Infection by a Giant Virus (AaV) Induces Widespread Physiological Reprogramming in Aureococcus anophagefferens CCMP1984 – A Harmful Bloom Algae Mohammad Moniruzzaman University of Tennessee, Knoxville Eric R. Gann University of Tennessee, Knoxville Steven W. Wilhelm University of Tennessee, Knoxville, [email protected] Follow this and additional works at: https://trace.tennessee.edu/utk_micrpubs Recommended Citation Moniruzzaman, Mohammad, Eric R. Gann, and Steven W. Wilhelm. “Infection by a Giant Virus (AaV) Induces Widespread Physiological Reprogramming in Aureococcus anophagefferens CCMP1984 – A Harmful Bloom Algae.” Frontiers in Microbiology 9 (2018). https://doi.org/10.3389/fmicb.2018.00752. This Article is brought to you for free and open access by the Microbiology at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Microbiology Publications and Other Works by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. fmicb-09-00752 April 18, 2018 Time: 16:59 # 1 ORIGINAL RESEARCH published: 19 April 2018 doi: 10.3389/fmicb.2018.00752 Infection by a Giant Virus (AaV) Induces Widespread Physiological Reprogramming in Aureococcus anophagefferens CCMP1984 – A Harmful Bloom Algae Mohammad Moniruzzaman1,2, Eric R. Gann1 and Steven W. Wilhelm1* 1 Department of Microbiology, The University of Tennessee, Knoxville, Knoxville, TN, United States, 2 Monterey Bay Aquarium Research Institute (MBARI), Moss Landing, CA, United States While viruses with distinct phylogenetic origins and different nucleic acid types can infect and lyse eukaryotic phytoplankton, “giant” dsDNA viruses have been found to Edited by: be associated with important ecological processes, including the collapse of algal Akio Adachi, Tokushima University, Japan blooms.
    [Show full text]
  • (10) Patent No.: US 8119385 B2
    US008119385B2 (12) United States Patent (10) Patent No.: US 8,119,385 B2 Mathur et al. (45) Date of Patent: Feb. 21, 2012 (54) NUCLEICACIDS AND PROTEINS AND (52) U.S. Cl. ........................................ 435/212:530/350 METHODS FOR MAKING AND USING THEMI (58) Field of Classification Search ........................ None (75) Inventors: Eric J. Mathur, San Diego, CA (US); See application file for complete search history. Cathy Chang, San Diego, CA (US) (56) References Cited (73) Assignee: BP Corporation North America Inc., Houston, TX (US) OTHER PUBLICATIONS c Mount, Bioinformatics, Cold Spring Harbor Press, Cold Spring Har (*) Notice: Subject to any disclaimer, the term of this bor New York, 2001, pp. 382-393.* patent is extended or adjusted under 35 Spencer et al., “Whole-Genome Sequence Variation among Multiple U.S.C. 154(b) by 689 days. Isolates of Pseudomonas aeruginosa” J. Bacteriol. (2003) 185: 1316 1325. (21) Appl. No.: 11/817,403 Database Sequence GenBank Accession No. BZ569932 Dec. 17. 1-1. 2002. (22) PCT Fled: Mar. 3, 2006 Omiecinski et al., “Epoxide Hydrolase-Polymorphism and role in (86). PCT No.: PCT/US2OO6/OOT642 toxicology” Toxicol. Lett. (2000) 1.12: 365-370. S371 (c)(1), * cited by examiner (2), (4) Date: May 7, 2008 Primary Examiner — James Martinell (87) PCT Pub. No.: WO2006/096527 (74) Attorney, Agent, or Firm — Kalim S. Fuzail PCT Pub. Date: Sep. 14, 2006 (57) ABSTRACT (65) Prior Publication Data The invention provides polypeptides, including enzymes, structural proteins and binding proteins, polynucleotides US 201O/OO11456A1 Jan. 14, 2010 encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides.
    [Show full text]
  • Biochemical and Transcriptome Analyses of a Novel Chlorophyll
    Wang et al. BMC Plant Biology 2014, 14:352 http://www.biomedcentral.com/1471-2229/14/352 RESEARCH ARTICLE Open Access Biochemical and transcriptome analyses of a novel chlorophyll-deficient chlorina tea plant cultivar Lu Wang1,2,3?,ChuanYue1,3?,HongliCao1,3, Yanhua Zhou1,3,JianmingZeng1,2,3,YajunYang1,2,3* and Xinchao Wang1,2,3* Abstract Background: The tea plant (Camellia sinensis (L.) O. Kuntze) is one of the most economically important woody crops. Recently, many leaf color genotypes have been developed during tea plant breeding and have become valuable materials in the processing of green tea. Although the physiological characteristics of some leaf color mutants of tea plants have been partially revealed, little is known about the molecular mechanisms leading to the chlorina phenotype in tea plants. Results: The yellow-leaf tea cultivar Zhonghuang 2 (ZH2) was selected during tea plant breeding. In comparison with Longjing 43 (LJ43), a widely planted green tea cultivar, ZH2 exhibited the chlorina phenotype and displayed significantly decreased chlorophyll contents. Transmission electron microscopy analysis revealed that the ultrastructure of the chloroplasts was disrupted, and the grana were poorly stacked in ZH2. Moreover, thecontentsoftheanineand free amino acids were significantly higher, whereas the contents of carotenoids, catechins and anthocyanin were lower in ZH2 than in LJ43. Microarray analysis showed that the expression of 259 genes related to amino acid metabolism, photosynthesis and pigment metabolism was significantly altered in ZH2 shoots compared with thoseofLJ43plants.Pathwayanalysisof4,902differentially expressed genes identified 24 pathways as being significantly regulated, including ?cysteine and methionine metabolism?, ?glycine, serine and threonine metabolism?, ?flavonoid biosynthesis?, ?porphyrin and chlorophyll metabolism?and ?carotenoid biosynthesis?.
    [Show full text]
  • Identification and Functional Characterization of the First Two
    Identification and functional characterization of the first two aromatic prenyltransferases implicated in the biosynthesis of furanocoumarins and prenylated coumarins in two plant families: Rutaceae and Apiaceae Fazeelat Karamat To cite this version: Fazeelat Karamat. Identification and functional characterization of the first two aromatic prenyl- transferases implicated in the biosynthesis of furanocoumarins and prenylated coumarins in two plant families: Rutaceae and Apiaceae. Agronomy. Université de Lorraine, 2013. English. NNT : 2013LORR0029. tel-01749560 HAL Id: tel-01749560 https://hal.univ-lorraine.fr/tel-01749560 Submitted on 29 Mar 2018 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. AVERTISSEMENT Ce document est le fruit d'un long travail approuvé par le jury de soutenance et mis à disposition de l'ensemble de la communauté universitaire élargie. Il est soumis à la propriété intellectuelle de l'auteur. Ceci implique une obligation de citation et de référencement lors de l’utilisation de ce document. D'autre part, toute contrefaçon, plagiat,
    [Show full text]
  • A Recruiting Protein of Geranylgeranyl Diphosphate Synthase Controls Metabolic Flux Toward Chlorophyll Biosynthesis in Rice
    A recruiting protein of geranylgeranyl diphosphate synthase controls metabolic flux toward chlorophyll biosynthesis in rice Fei Zhoua,1, Cheng-Yuan Wangb,1, Michael Gutensohnc,1, Ling Jiangd, Peng Zhangb, Dabing Zhange, Natalia Dudarevaf,2, and Shan Lua,2 aState Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China; bState Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China; cDivision of Plant and Soil Sciences, West Virginia University, Morgantown, WV 26505; dState Key Laboratory of Crop Genetics and Germplasm Enhancement, Research Center of Jiangsu Plant Gene Engineering, Nanjing Agricultural University, Nanjing 210095, China; eState Key Laboratory of Hybrid Rice, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China; and fDepartment of Biochemistry, Purdue University, West Lafayette, IN 47907 Edited by Joanne Chory, The Salk Institute for Biological Studies and Howard Hughes Medical Institute, La Jolla, CA, and approved May 18, 2017 (received for review April 6, 2017) In plants, geranylgeranyl diphosphate (GGPP) is produced by is shared by several vital metabolic pathways, particularly in plastidic GGPP synthase (GGPPS) and serves as a precursor for vital plastids, including the biosynthesis of carotenoids and their deriv- metabolic branches, including chlorophyll, carotenoid, and gibber- atives (e.g., plant hormones abscisic acid and strigolactones), as ellin biosynthesis. However, molecular mechanisms regulating GGPP well as gibberellins, plastoquinones, tocopherols, and the phytyl tail allocation among these biosynthetic pathways localized in the same of chlorophylls (3). However, it remains largely unknown how al- subcellular compartment are largely unknown.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8,535,916 B2 Del Cardayre Et Al
    US008535916B2 (12) United States Patent (10) Patent No.: US 8,535,916 B2 Del Cardayre et al. (45) Date of Patent: Sep. 17, 2013 (54) MODIFIEDMICROORGANISMS AND USES (51) Int. Cl. THEREFOR CI2P 7/64 (2006.01) (52) U.S. Cl. (75) Inventors: Stephen B. Del Cardayre, Belmont, CA USPC .......................................................... 435/134 (US); Shane Brubaker, Oakland, CA (58) Field of Classification Search (US); Jay D. Keasling, Berkeley, CA USPC .......................................................... 435/134 (US) See application file for complete search history. (73) Assignee: LS9, Inc., South San Francisco, CA (US) (56) References Cited (*) Notice: Subject to any disclaimer, the term of this PUBLICATIONS patent is extended or adjusted under 35 IPER PCT/US2007/003736 (2007).* U.S.C. 154(b) by 497 days. Ohmiya et al. Journal of Bioscience and Bioengineering,95(6): 549 561 (2003).* (21) Appl. No.: 12/526,209 * cited by examiner (22) PCT Filed: Feb. 13, 2007 Primary Examiner — Tekchand Saidha (86). PCT No.: PCT/US2007/003736 (74) Attorney, Agent, or Firm — Brigitte A. Hajos S371 (c)(1), (57) ABSTRACT (2), (4) Date: Dec. 6, 2010 The invention provides a genetically modified microorgan ism that acquires the ability to consume a renewable feed (87) PCT Pub. No.: WO2008/100251 stock (such as cellulose) and produce products. This organ PCT Pub. Date: Aug. 21, 2008 ism can be used to ferment cellulose, one of the most abundant renewable resources available, and produce prod (65) Prior Publication Data uctS. US 2011 FOO97769 A1 Apr. 28, 2011 8 Claims, 1 Drawing Sheet U.S. Patent Sep. 17, 2013 US 8,535,916 B2 Anchor Peptide - Scaffoldin US 8,535,916 B2 1.
    [Show full text]
  • A Mutant Allele of Ζ-Carotene Isomerase (Z-ISO)
    Rodrigo et al. BMC Plant Biology (2019) 19:465 https://doi.org/10.1186/s12870-019-2078-2 RESEARCH ARTICLE Open Access A mutant allele of ζ-carotene isomerase (Z- ISO) is associated with the yellow pigmentation of the “Pinalate” sweet orange mutant and reveals new insights into its role in fruit carotenogenesis María J. Rodrigo1* , Joanna Lado1,2, Enriqueta Alós1, Berta Alquézar1,3, Orly Dery4, Joseph Hirschberg4 and Lorenzo Zacarías1 Abstract Background: Fruit coloration is one of the main quality parameters of Citrus fruit primarily determined by genetic factors. The fruit of ordinary sweet orange (Citrus sinensis) displays a pleasant orange tint due to accumulation of carotenoids, representing β,β-xanthophylls more than 80% of the total content. ‘Pinalate’ is a spontaneous bud mutant, or somatic mutation, derived from sweet orange ‘Navelate’, characterized by yellow fruits due to elevated proportions of upstream carotenes and reduced β,β-xanthophylls, which suggests a biosynthetic blockage at early steps of the carotenoid pathway. Results: To identify the molecular basis of ‘Pinalate’ yellow fruit, a complete characterization of carotenoids profile together with transcriptional changes in carotenoid biosynthetic genes were performed in mutant and parental fruits during development and ripening. ‘Pinalate’ fruit showed a distinctive carotenoid profile at all ripening stages, accumulating phytoene, phytofluene and unusual proportions of 9,15,9′-tri-cis- and 9,9′-di-cis-ζ-carotene, while content of downstream carotenoids was significantly decreased. Transcript levels for most of the carotenoid biosynthetic genes showed no alterations in ‘Pinalate’; however, the steady-state level mRNA of ζ-carotene isomerase (Z-ISO), which catalyses the conversion of 9,15,9′-tri-cis- to 9,9′-di-cis-ζ-carotene, was significantly reduced both in ‘Pinalate’ fruit and leaf tissues.
    [Show full text]