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Fatty alcohol production in engineered E. coli expressing Marinobacter fatty acyl-CoA reductases

  • Bioenergy and biofuels
  • Published:
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Abstract

Although successful production of fatty alcohols in metabolically engineered Escherichia coli with heterologous expression of fatty acyl-CoA reductase has been reported, low biosynthetic efficiency is still a hurdle to be overcome. In this study, we examined the characteristics of two fatty acyl-CoA reductases encoded by Maqu_2220 and Maqu_2507 genes from Marinobacter aquaeolei VT8 on fatty alcohol production in E. coli. Fatty alcohols with diversified carbon chain length were obtained by co-expressing Maqu_2220 with different carbon chain length-specific acyl-ACP thioesterases. Both fatty acyl-CoA reductases displayed broad substrate specificities for C12–C18 fatty acyl chains in vivo. The optimized mutant strain of E. coli carrying the modified tesA gene and fadD gene from E. coli and Maqu_2220 gene from Marinobacter aquaeolei VT8 produced fatty alcohols at a remarkable level of 1.725 g/L under the fermentation condition.

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References

  • Black PN, DiRusso CC, Metzger AK, Heimert TL (1992) Cloning, sequencing, and expression of the fadD gene of Escherichia coli encoding acyl coenzyme A synthetase. J Biol Chem 267:25513–25520

    PubMed  CAS  Google Scholar 

  • Carlsson AS, Yilmaz JL, Green AG, Stymne S, Hofvander P (2011) Replacing fossil oil with fresh oil - with what and for what? Eur J Lipid Sci Technol 113:812–831

    Article  PubMed  CAS  Google Scholar 

  • Cheng JB (2004) Mammalian wax biosynthesis: I. Identification of two fatty acyl-coenzyme A reductases with different substrate specificities and tissue distributions. J Biol Chem 279:37789–37797

    Article  PubMed  CAS  Google Scholar 

  • Cho H, Cronan JE Jr (1995) Defective export of a periplasmic enzyme disrupts regulation of fatty acid synthesis. J Biol Chem 270:4216–4219

    Article  PubMed  CAS  Google Scholar 

  • Davis MS (2000) Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J Biol Chem 275:28593–28598

    Article  PubMed  CAS  Google Scholar 

  • Doan TTP, Carlsson AS, Hamberg M, Bülow L, Stymne S, Olsson P (2009) Functional expression of five Arabidopsis fatty acyl-CoA reductase genes in Escherichia coli. J Plant Physiol 166:787–796

    Article  PubMed  CAS  Google Scholar 

  • Domergue F, Vishwanath SJ, Joubes J, Ono J, Lee JA, Bourdon M, Alhattab R, Lowe C, Pascal S, Lessire R, Rowland O (2010) Three Arabidopsis fatty acyl-Coenzyme A reductases, FAR1, FAR4, and FAR5, generate primary fatty alcohols associated with suberin deposition. Plant Physiol 153:1539–1554

    Article  PubMed  CAS  Google Scholar 

  • Duan Y, Zhu Z, Cai K, Tan X, Lu X (2011) De novo biosynthesis of biodiesel by Escherichia coli in optimized fed-batch cultivation. PLoS One 6:e20265

    Article  PubMed  CAS  Google Scholar 

  • Fortman JL, Chhabra S, Mukhopadhyay A, Chou H, Lee TS, Steen E, Keasling JD (2008) Biofuel alternatives to ethanol: pumping the microbial well. Trends Biotechnol 26:375–381

    Article  PubMed  CAS  Google Scholar 

  • Gervajio GC (2005) Fatty acids and derivatives from coconut oil. Bailey’s Industrial Oil and Fat Products. John Wiley & Sons, Inc

    Google Scholar 

  • Hofvander P, Doan TTP, Hamberg M (2011) A prokaryotic acyl-CoA reductase performing reduction of fatty acyl-CoA to fatty alcohol. FEBS Lett 585:3538–3543

    Article  PubMed  CAS  Google Scholar 

  • Jiang P, Cronan JE Jr (1994) Inhibition of fatty acid synthesis in Escherichia coli in the absence of phospholipid synthesis and release of inhibition by thioesterase action. J Bacteriol 176:2814–2821

    PubMed  CAS  Google Scholar 

  • Kaczmarzyk D, Fulda M (2010) Fatty acid activation in cyanobacteria mediated by acyl-acyl carrier protein synthetase enables fatty acid recycling. Plant Physiol 152:1598–1610

    Article  PubMed  CAS  Google Scholar 

  • Kalscheuer R, Stolting T, Steinbuchel A (2006) Microdiesel: Escherichia coli engineered for fuel production. Microbiology 152:2529–2536

    Article  PubMed  CAS  Google Scholar 

  • Kameda K, Nunn WD (1981) Purification and characterization of acyl coenzyme A synthetase from Escherichia coli. J Biol Chem 256:5702–5707

    PubMed  CAS  Google Scholar 

  • Kanya TCS, Rao LJ, Sastry MCS (2007) Characterization of wax esters, free fatty alcohols and free fatty acids of crude wax from sunflower seed oil refineries. Food Chem 101:1552–1557

    Article  CAS  Google Scholar 

  • Li M, Zhang X, Agrawal A, San K-Y (2012) Effect of acetate formation pathway and long chain fatty acid CoA-ligase on the free fatty acid production in E. coli expressing acy-ACP thioesterase from Ricinus communis. Metab Eng 14:380–387

    Article  PubMed  CAS  Google Scholar 

  • Liu T, Vora H, Khosla C (2010) Quantitative analysis and engineering of fatty acid biosynthesis in E. coli. Metab Eng 12:378–386

    Article  PubMed  CAS  Google Scholar 

  • Liu XSJ, Curtiss R (2011) Fatty acid production in genetically modified cyanobacteria. Proc Natl Acad Sci USA 6899–6904

  • Lu X, Vora H, Khosla C (2008) Overproduction of free fatty acids in E. coli: implications for biodiesel production. Metab Eng 10:333–339

    Article  PubMed  CAS  Google Scholar 

  • Metz JG, Pollard MR, Anderson L, Hayes TR, Lassner MW (2000) Purification of a jojoba embryo fatty acyl-coenzyme A reductase and expression of its cDNA in high erucic acid rapeseed. Plant Physiol 122:635–644

    Article  PubMed  CAS  Google Scholar 

  • Peralta-Yahya PP, Zhang F, del Cardayre SB, Keasling JD (2012) Microbial engineering for the production of advanced biofuels. Nature 488:320–328

    Article  PubMed  CAS  Google Scholar 

  • 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 179:2969–2975

    PubMed  CAS  Google Scholar 

  • Rizzo WB, Craft DA (2000) Sjögren–Larsson syndrome: accumulation of free fatty alcohols in cultured fibroblasts and plasma. J Lipid Res 41:1077–1081

    PubMed  CAS  Google Scholar 

  • Rowland O, Zheng H, Hepworth SR, Lam P, Jetter R, Kunst L (2006) CER4 encodes an alcohol-forming fatty acyl-coenzyme A reductase involved in cuticular wax production in Arabidopsis. Plant Physiol 142:866–877

    Article  PubMed  CAS  Google Scholar 

  • Schirmer A, Rude MA, Li X, Popova E, del Cardayre SB (2010) Microbial biosynthesis of alkanes. Science 329:559–562

    Article  PubMed  CAS  Google Scholar 

  • Steen EJ, Kang Y, Bokinsky G, Hu Z, Schirmer A, McClure A, del Cardayre SB, Keasling JD (2010) Microbial production of fatty-acid-derived fuels and chemicals from plant biomass. Nature 463:559–562

    Article  PubMed  CAS  Google Scholar 

  • Tan X, Yao L, Gao Q, Wang W, Qi F, Lu X (2011) Photosynthesis driven conversion of carbon dioxide to fatty alcohols and hydrocarbons in cyanobacteria. Metab Eng 13:169–176

    Article  PubMed  CAS  Google Scholar 

  • Teerawanichpan P, Qiu X (2010) Fatty acyl-CoA reductase and wax synthase from Euglena gracilis in the biosynthesis of medium-chain wax esters. Lipids 45:263–273

    Article  PubMed  CAS  Google Scholar 

  • Teerawanichpan P, Robertson AJ, Qiu X (2010) A fatty acyl-CoA reductase highly expressed in the head of honey bee (Apis mellifera) involves biosynthesis of a wide range of aliphatic fatty alcohols. Insect Biochem Mol Biol 40:641–649

    Article  PubMed  CAS  Google Scholar 

  • Vioque J, Kolattukudy PE (1997) Resolution and purification of an aldehyde-generating and an alcohol-generating fatty acyl-CoA reductase from pea leaves (Pisum sativum L.). Arch Biochem Biophys 340:64–72

    Article  PubMed  CAS  Google Scholar 

  • Voelker TA, Davies HM (1994) Alteration of the specificity and regulation of fatty acid synthesis of Escherichia coli by expression of a plant medium-chain acyl-acyl carrier protein thioesterase. J Bacteriol 176:7320–7327

    PubMed  CAS  Google Scholar 

  • Wahlen BD, Oswald WS, Seefeldt LC, Barney BM (2009) Purification, characterization, and potential bacterial wax production role of an NADPH-dependent fatty aldehyde reductase from Marinobacter aquaeolei VT8. Appl Environ Microbiol 75:2758–2764

    Article  PubMed  CAS  Google Scholar 

  • Willis RM, Wahlen BD, Seefeldt LC, Barney BM (2011) Characterization of a fatty acyl-CoA reductase from Marinobacter aquaeolei VT8: a bacterial enzyme catalyzing the reduction of fatty acyl-CoA to fatty alcohol. Biochemistry 50:10550–10558

    Article  PubMed  CAS  Google Scholar 

  • Yuan L, Voelker TA, Hawkins DJ (1995) Modification of the substrate specificity of an acyl-acyl carrier protein thioesterase by protein engineering. Proc Natl Acad Sci USA 92:10639–10643

    Article  PubMed  CAS  Google Scholar 

  • 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 14:653–660

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Li M, Agrawal A, San K-Y (2011) Efficient free fatty acid production in Escherichia coli using plant acyl-ACP thioesterases. Metab Eng 13:713–722

    Article  PubMed  CAS  Google Scholar 

  • 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 Fact 11:65

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by “100-Talent Program of the Chinese Academy of Sciences” foundation (Grant O91001110A).

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Authors and Affiliations

  1. Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China

    Aiqiu Liu, Xiaoming Tan, Lun Yao & Xuefeng Lu

  2. Ocean College, Shandong University (Weihai), Weihai, 264209, China

    Aiqiu Liu

  3. University of Chinese Academy of Sciences, Beijing, 100049, China

    Aiqiu Liu & Lun Yao

Authors
  1. Aiqiu Liu
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  2. Xiaoming Tan
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  3. Lun Yao
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  4. Xuefeng Lu
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Correspondence to Xuefeng Lu.

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Liu, A., Tan, X., Yao, L. et al. Fatty alcohol production in engineered E. coli expressing Marinobacter fatty acyl-CoA reductases. Appl Microbiol Biotechnol 97, 7061–7071 (2013). https://doi.org/10.1007/s00253-013-5027-2

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  • DOI: https://doi.org/10.1007/s00253-013-5027-2

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