Improving alkane synthesis in Escherichia coli via metabolic engineering

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• 作者：Xuejiao Song ; Haiying Yu ; Kun Zhu
• 关键词：Alkane biosynthesis ; Biofuels ; Fatty acid biosynthesis ; Aldehyde reductase ; fadR ; Escherichia coli
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：100
• 期：2
• 页码：757-767
• 全文大小：706 KB
• 参考文献：Akhtar MK, Turner NJ, Jones PR (2013) Carboxylic acid reductase is a versatile enzyme for the conversion of fatty acids into fuels and chemical commodities. Proc Natl Acad Sci U S A 110(1):87–92PubMed PubMedCentral CrossRef
  Andre C, Kim SW, Yu XH, Shanklin J (2013) Fusing catalase to an alkane-producing enzyme maintains enzymatic activity by converting the inhibitory byproduct H2O2 to the cosubstrate O2. Proc Natl Acad Sci U S A 110(8):3191–3196PubMed PubMedCentral CrossRef
  Atsumi S, Wu TY, Eckl EM, Hawkins SD, Buelter T, Liao JC (2010) Engineering the isobutanol biosynthetic pathway in Escherichia coli by comparison of three aldehyde reductase/alcohol dehydrogenase genes. Appl Microbiol Biotechnol 85(3):651–657PubMed PubMedCentral CrossRef
  Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2:2006.0008PubMed PubMedCentral CrossRef
  Beller HR, Goh EB, Keasling JD (2010) Genes involved in long-chain alkene biosynthesis in Micrococcus luteus. Appl Environ Microbiol 76(4):1212–1223
  Bernard A, Domergue F, Pascal S, Jetter R, Renne C, Faure J-D, Haslam RP, Napier JA, Lessire R, Joubès J (2012) Reconstitution of plant alkane biosynthesis in yeast demonstrates that Arabidopsis ECERIFERUM1 and ECERIFERUM3 are core components of a very-long-chain alkane synthesis complex. Plant Cell 24(7):3106–3118
  Cherepanov PP, Wackernagel W (1995) Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158(1):9–14PubMed CrossRef
  Choi YJ, Lee SY (2013) Microbial production of short-chain alkanes. Nature 502(7472):571–574PubMed CrossRef
  Cronan JE (1997) In vivo evidence that acyl coenzyme A regulates DNA binding by the Escherichia coli FadR global transcription factor. J Bacteriol 179(5):1819–1823PubMed PubMedCentral
  Dellomonaco C, Clomburg JM, Miller EN, Gonzalez R (2011) Engineered reversal of the β-oxidation cycle for the synthesis of fuels and chemicals. Nature 476(7360):355–359PubMed CrossRef
  Fujita Y, Matsuoka H, Hirooka K (2007) Regulation of fatty acid metabolism in bacteria. Mol Microbiol 66(4):829–839PubMed CrossRef
  Harger M, Zheng L, Moon A, Ager C, An JH, Choe C, Lai YL, Mo B, Zong D, Smith MD, Egbert RG, Mills JH, Baker D, Pultz IS, Siegel JB (2013) Expanding the product profile of a microbial alkane biosynthetic pathway. ACS Synth Biol 2(1):59–62PubMed CrossRef
  Howard TP, Middelhaufe S, Moore K, Edner C, Kolak DM, Taylor GN, Parker DA, Lee R, Smirnoff N, Aves SJ, Love J (2013) Synthesis of customized petroleum-replica fuel molecules by targeted modification of free fatty acid pools in Escherichia coli. Proc Natl Acad Sci U S A 110(19):7636–7641PubMed PubMedCentral CrossRef
  Janßen HJ, Steinbüchel A (2014) Fatty acid synthesis in Escherichia coli and its applications towards the production of fatty acid based biofuels. Biotechnol Biofuels 7:7PubMed PubMedCentral CrossRef
  Kass LR, Bloch K (1967) On the enzymatic synthesis of unsaturated fatty acids in Escherichia coli. Proc Natl Acad Sci U S A 58(3):1168–1173PubMed PubMedCentral CrossRef
  Knothe G (2009) Improving biodiesel fuel properties by modifying fatty ester composition. Energy Environ Sci 2(7):759–766CrossRef
  Lennen RM, Braden DJ, West RA, Dumesic JA, Pfleger BF (2010) A process for microbial hydrocarbon synthesis: overproduction of fatty acids in Escherichia coli and catalytic conversion to alkanes. Biotechnol Bioeng 106(2):193–202PubMed CrossRef
  Liu A, Zhu T, Lu X, Song L (2013) Hydrocarbon profiles and phylogenetic analyses of diversified cyanobacterial species. Appl Energy 111:383–393CrossRef
  Liu X, Yu H, Jiang X, Ai G, Yu B, Zhu K (2015) Biosynthesis of butenoic acid through fatty acid biosynthesis pathway in Escherichia coli. Appl Microbiol Biotechnol 99(4):1795–1804PubMed CrossRef
  Marrakchi H, Choi K-H, Rock CO (2002) A new mechanism for anaerobic unsaturated fatty acid formation in Streptococcus pneumoniae. J Biol Chem 277(47):44809–44816PubMed CrossRef
  Mendez-Perez D, Begemann MB, Pfleger BF (2011) Modular synthase-encoding gene involved in alpha-olefin biosynthesis in Synechococcus sp. strain PCC 7002. Appl Environ Microbiol 77(12):4264–4267PubMed PubMedCentral CrossRef
  Nunn WD, Giffin K, Clark D, Cronan JE (1983) Role for fadR in unsaturated fatty acid biosynthesis in Escherichia coli. J Bacteriol 154(2):554–560PubMed PubMedCentral
  Peralta-Yahya PP, Zhang F, del Cardayre SB, Keasling JD (2012) Microbial engineering for the production of advanced biofuels. Nature 488(7411):320–328PubMed CrossRef
  Perez JM, Arenas FA, Pradenas GA, Sandoval JM, Vasquez CC (2008) Escherichia coli YqhD exhibits aldehyde reductase activity and protects from the harmful effect of lipid peroxidation-derived aldehydes. J Biol Chem 283(12):7346–7353PubMed CrossRef
  Rahman Z, Sung BH, Yi J-Y, Bui LM, Lee JH, Kim SC (2014) Enhanced production of n-alkanes in Escherichia coli by spatial organization of biosynthetic pathway enzymes. J Biotechnol 192:187–191CrossRef
  Reiser S, Somerville C (1997) Isolation of mutants of Acinetobacter calcoaceticus deficient in wax ester synthesis and complementation of one mutation with a gene encoding a fatty acyl coenzyme A reductase. J Bacteriol 179(9):2969–2975PubMed PubMedCentral
  Rodriguez GM, Atsumi S (2012) Isobutyraldehyde production from Escherichia coli by removing aldehyde reductase activity. Microb Cell Factories 11:90CrossRef
  Rodriguez GM, Atsumi S (2014) Toward aldehyde and alkane production by removing aldehyde reductase activity in Escherichia coli. Metab Eng 25:227–237PubMed PubMedCentral CrossRef
  Rude MA, Baron TS, Brubaker S, Alibhai M, Del Cardayre SB, Schirmer A (2011) Terminal olefin (1-alkene) biosynthesis by a novel p450 fatty acid decarboxylase from Jeotgalicoccus species. Appl Environ Microbiol 77(5):1718–1727PubMed PubMedCentral CrossRef
  Schirmer A, Rude MA, Li X, Popova E, del Cardayre SB (2010) Microbial biosynthesis of alkanes. Science 329(5991):559–562PubMed CrossRef
  Sukovich DJ, Seffernick JL, Richman JE, Gralnick JA, Wackett LP (2010) Widespread head-to-head hydrocarbon biosynthesis in bacteria and role of OleA. Appl Environ Microbiol 76(12):3850–3862PubMed PubMedCentral CrossRef
  Sulzenbacher G, Alvarez K, van den Heuvel RHH, Versluis C, Spinelli S, Campanacci V, Valencia C, Cambillau C, Eklund H, Tegoni M (2004) Crystal structure of Escherichia coli alcohol dehydrogenase YqhD: evidence of a covalently modified NADP coenzyme. J Mol Biol 342(2):489–502PubMed CrossRef
  Thomason LC, Costantino N, Court DL (2007) E. coli genome manipulation by P1 transduction. Curr Protoc Mol Biol 79:1.17.11–11.17.18
  Tsay JT, Oh W, Larson TJ, Jackowski S, Rock CO (1992) Isolation and characterization of the beta-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12. J Biol Chem 267(10):6807–6814PubMed
  Xu P, Vansiri A, Bhan N, Koffas MAG (2012) ePathBrick: a synthetic biology platform for engineering metabolic pathways in E. coli. ACS Synth Biol 1(7):256–266PubMed CrossRef
  Zhang F, Ouellet M, Batth TS, Adams PD, Petzold CJ, Mukhopadhyay A, Keasling JD (2012) Enhancing fatty acid production by the expression of the regulatory transcription factor FadR. Metab Eng 14(6):653–660PubMed CrossRef
  Zhang J, Lu X, Li J-J (2013) Conversion of fatty aldehydes into alk (a/e)nes by in vitro reconstituted cyanobacterial aldehyde-deformylating oxygenase with the cognate electron transfer system. Biotechnol Biofuels 6(1):1–10PubMed PubMedCentral CrossRef
  Zheng YN, Li LL, Liu Q, Yang JM, Wang XW, Liu W, Xu X, Liu H, Zhao G, Xian M (2012) Optimization of fatty alcohol biosynthesis pathway for selectively enhanced production of C12/14 and C16/18 fatty alcohols in engineered Escherichia coli. Microb Cell Factories 11:65CrossRef
  Zhu K, Zhang Y-M, Rock CO (2009) Transcriptional regulation of membrane lipid homeostasis in Escherichia coli. J Biol Chem 284(50):34880–34888PubMed PubMedCentral CrossRef
  
• 作者单位：Xuejiao Song (1) (2)
  Haiying Yu (1)
  Kun Zhu (1)
  
  1. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China
  2. University of Chinese Academy of Sciences, Beijing, 100049, China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Concerns about energy security and global petroleum supply have made the production of renewable biofuels an industrial imperative. The ideal biofuels are n-alkanes in that they are chemically and structurally identical to the fossil fuels and can “drop in” to the transportation infrastructure. In this work, an Escherichia coli strain that produces n-alkanes was constructed by heterologous expression of acyl-acyl carrier protein (ACP) reductase (AAR) and aldehyde deformylating oxygenase (ADO) from Synechococcus elongatus PCC7942. The accumulation of alkanes ranged from 3.1 to 24.0 mg/L using different expressing strategies. Deletion of yqhD, an inherent aldehyde reductase in E. coli, or overexpression of fadR, an activator for fatty acid biosynthesis, exhibited a nearly twofold increase in alkane titers, respectively. Combining yqhD deletion and fadR overexpression resulted in a production titer of 255.6 mg/L in E. coli, and heptadecene was the most abundant product. Keywords Alkane biosynthesis Biofuels Fatty acid biosynthesis Aldehyde reductase fadR Escherichia coli