Abstract
Butenoic acid is a C4 short-chain unsaturated fatty acid mainly used in the preparation of resins, pharmaceuticals, and fine chemicals. However, butenoic acid derived from petroleum is costly and unfriendly to the environment. Here, we report a novel biosynthetic strategy to produce butenoic acid by utilizing the intermediate of fatty acid biosynthesis pathway in engineered Escherichia coli. A thioesterase gene (B. thetaiotaomicron thioesterase (bTE)) from Bacteroides thetaiotaomicron was heterologously expressed in E. coli to specifically convert butenoyl-acyl carrier protein (ACP), a fatty acid biosynthesis intermediate, to butenoic acid. The titer of butenoic acid ranged from 0.07 to 11.4 mg/L in four different E. coli strains with varied expressing vectors. Deletion of endogenous fadD gene (encoding acyl-CoA synthetase) to block fatty acid oxidation improved the butenoic acid production in all strains to some extent. The highest butenoic acid accumulation of 18.7 mg/L was obtained in strain XP-2 (BL21-∆fadD/pET28a-bTE). Moreover, partially inhibiting the enoyl-ACP reductase (FabI) of strain XP-2 by triclosan increased butenoic acid production by threefold, and the butenoic acid titer was further increased to 161.4 mg/L by supplying glucose and tryptone in the M9 medium. Fed-batch fermentation of this strain further enhanced butenoic acid production to 4.0 g/L within 48 h. The butenoic acid tolerance assay revealed that this strain could tolerate 15–20 g/L of butenoic acid.
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References
Abdel-Rahman MA, Tashiro Y, Sonomoto K (2013) Recent advances in lactic acid production by microbial fermentation processes. Biotechnol Adv 31:877–902
Ai GM, Zhu JX, Dong XZ, Sun T (2013) Simultaneous characterization of methane and carbon dioxide produced by cultured methanogens using gas chromatography/isotope ratio mass spectrometry and gas chromatography/mass spectrometry. Rapid Commun Mass Spectrom 27:1935–1944
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 2:2006.0008
Barnes EM (1975) Long chain fatty acyl-thioesterases I and II from Escherichia coli. Meth Enzymol 35:102–109
Barnes EM, Swindell A, Wakil SJ (1970) Purification and properties of a palmityl thioesterase II from Escherichia coli. J Biol Chem 245:3122–3128
Black PN, DiRusso C, Metzger A, Heimert T (1992) Cloning, sequencing, and expression of the fadD gene of Escherichia coli encoding acyl coenzyme A synthetase. J Biol Chem 267:25513–25520
Cantu DC, Chen Y, Reilly PJ (2010) Thioesterases: a new perspective based on their primary and tertiary structures. Protein Sci 19:1281–1295
Cho H, Cronan J (1993) Escherichia coli thioesterase I, molecular cloning and sequencing of the structural gene and identification as a periplasmic enzyme. J Biol Chem 268:9238–9245
Cronan JE, Thomas J (2009) Bacterial fatty acid synthesis and its relationships with polyketide synthetic pathways. Methods Enzymol 459:395–433
Cummings J, Pomare E, Branch W, Naylor C, Macfarlane G (1987) Short chain fatty acids in human large intestine, portal, hepatic and venous blood. Gut 28:1221–1227
Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645
Davis MS, Solbiati J, Cronan JE (2000) Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J Biol Chem 275:28593–28598
Dehesh K, Edwards P, Hayes T, Cranmer AM, Fillatti J (1996) Two novel thioesterases are key determinants of the bimodal distribution of acyl chain length of Cuphea palustris seed oil. Plant Physiol 110:203–210
Dellomonaco C, Clomburg JM, Miller EN, Gonzalez R (2011) Engineered reversal of the β-oxidation cycle for the synthesis of fuels and chemicals. Nature 476:355–359
Escalada MG, Harwood JL, Maillard JY, Ochs D (2005) Triclosan inhibition of fatty acid synthesis and its effect on growth of Escherichia coli and Pseudomonas aeruginosa. J Antimicrob Chemother 55:879–882
Fujita Y, Matsuoka H, Hirooka K (2007) Regulation of fatty acid metabolism in bacteria. Mol Microbiol 66:829–839
Ghanta M, Fahey D, Subramaniam B (2013) Environmental impacts of ethylene production from diverse feedstocks and energy sources. Appl Petrochem Res 4:167–179
Handke P, Lynch SA, Gill RT (2011) Application and engineering of fatty acid biosynthesis in Escherichia coli for advanced fuels and chemicals. Metab Eng 13:28–37
He L, Xu YQ, Zhang XH (2008) Medium factor optimization and fermentation kinetics for phenazine-1-carboxylic acid production by Pseudomonas sp M18G. Biotechnol Bioeng 100:250–259
Heath RJ, Rock CO (2000) Microbiology: a triclosan-resistant bacterial enzyme. Nature 406:145–146
Hu G, Li J, Zeng G (2013) Recent development in the treatment of oily sludge from petroleum industry: a review. J Hazard Mater 261:470–490
Jang M, Kamens RM (2001) Characterization of secondary aerosol from the photooxidation of toluene in the presence of NOx and 1-propene. Environ Sci Technol 35:3626–3639
Jeffries TW (2005) Ethanol fermentation on the move. Nat Biotechnol 23:40–41
Jensen KF (1993) The Escherichia coli K-12 “wild types” W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol 175:3401–3407
Jing FY, Cantu D, Tvaruzkova J, Chipman J, Nikolau B, Yandeau-Nelson M, Reilly P (2011) Phylogenetic and experimental characterization of an acyl-ACP thioesterase family reveals significant diversity in enzymatic specificity and activity. BMC Biochem 12:44
Kang Y, Durfee T, Glasner JD, Qiu Y, Frisch D, Winterberg KM, Blattner FR (2004) Systematic mutagenesis of the Escherichia coli genome. J Bacteriol 186:4921–4930
Lan EI, Liao JC (2013) Microbial synthesis of n-butanol, isobutanol, and other higher alcohols from diverse resources. Bioresour Technol 135:339–349
Li J, Zhao JB, Zhao M, Yang YL, Jiang WH, Yang S (2010) Screening and characterization of butanol-tolerant micro-organisms. Lett Appl Microbiol 50:373–379
Lofty WA, Ghanem KM, El-Helow ER (2007) Citric acid production by a novel Aspergillus niger isolate: II. Optimization of process parameters through statistical experimental designs. Bioresour Technol 98:3470–3477
Lu H, Tonge PJ (2008) Inhibitors of FabI, an enzyme drug target in the bacterial fatty acid biosynthesis pathway. Acc Chem Res 41:11–20
Ma R, Ouyang J, Li X, Lian ZN, Cai C (2012) Simultaneous determination of organic acids and saccharides in lactic acid fermentation broth from biomass using high performance liquid chromatography. Se Pu 30:62–66
Masih M, Algahtani I, De Mello L (2010) Price dynamics of crude oil and the regional ethylene markets. Energy Econ 32:1435–1444
McMurry LM, Oethinger M, Levy SB (1998) Triclosan targets lipid synthesis. Nature 394:531–532
Nikolau BJ, Perera MA, Brachova L, Shanks B (2008) Platform biochemicals for a biorenewable chemical industry. Plant J Cell Mol Biol 54:536–545
Reitzer L (2003) Nitrogen assimilation and global regulation in Escherichia coli. Ann Rev Microbiol 57:155–176
Riesenberg D, Menzel K, Schulz V, Schumann K, Veith G, Zuber G, Knorre WA (1990) High cell density fermentation of recombinant Escherichia coli expressing human interferon alpha 1. Appl Microbiol Biotechnol 34:77–82
Rollat I, Samain H, Morel O (2006) Reshapable hair styling composition comprising (meth)acrylic copolymers of four or more monomers. US 07122175
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Saraydin D, Karadag E, Guven O (1998) The releases of agrochemicals from radiation induced acrylamide crotonic acid hydrogels. Polym Bull 41:577–584
Schneider J, Niermann K, Wendisch VF (2011) Production of the amino acids L-glutamate, L-lysine, L-ornithine and L-arginine from arabinose by recombinant Corynebacterium glutamicum. J Biotechnol 154:191–198
Schulz RP, Blumenstein J, Kohlpaintner C (2000) Crotonaldehyde and crotonic acid. Ullmann’s encyclopedia of chemical technology. Wiley, Weinheim
Schummer C, Delhomme O, Appenzeller BMR, Wennig R, Millet M (2009) Comparison of MTBSTFA and BSTFA in derivatization reactions of polar compounds prior to GC/MS analysis. Talanta 77:1473–1482
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
Torella JP, Ford TJ, Kim SN, Chen AM, Way JC, Silver PA (2013) Tailored fatty acid synthesis via dynamic control of fatty acid elongation. Proc Natl Acad Sci U S A 110:11290–11295
Tsay J-T, Oh W, Larson T, Jackowski S, Rock C (1992) Isolation and characterization of the beta-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12. J Biol Chem 267:6807–6814
Vanderheide E, Zwinkels M, Gerritsen A, Scholten J (1992) Oxidation of ethylene to acetaldehyde over a heterogenized surface-vanadate wacker catalyst in the absence of gaseous oxygen. Appl Catal A Gen 86:181–198
Verwoert I, Verbree E, Van der Linden K, Nijkamp H, Stuitje A (1992) Cloning, nucleotide sequence, and expression of the Escherichia coli fabD gene, encoding malonyl coenzyme A-acyl carrier protein transacylase. J Bacteriol 174:2851–2857
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
Wakaki S, Yamamoto T, Enoki H (2008) Stabilizing agent for chlorine containing polymer used for chlorine containing polymer composition, contains epoxy-group containing acrylic resin, amino crotonic-acid ester, polyhydric alcohol and/or hindered amine or phenyl indole. WO2008087784-A1; JP2008195912-A; JP5192182-B2
Wang Z, Yan M, Chen X, Li DS, Qin L, Li ZJ, Yao J, Liang XL (2013) Mixed culture of Saccharomyces cerevisiae and Acetobacter pasteurianus for acetic acid production. Biochem Eng J 79:41–45
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 U S A 92:10639–10643
Zeiller J-J, Dumas H, Guyard-Dangremont V, Berard I, Contard F, Guerrier D, Ferrand G, Bonhomme Y (2012) Butenoic acid derivatives, processes for the preparation thereof, pharmaceutical compositions comprising them, and use for the treatment of dyslipidaemia, atherosclerosis and diabetes. US 08247448
Zhang XJ, Li M, Agrawal A, San KY (2011) Efficient free fatty acid production in Escherichia coli using plant acyl-ACP thioesterases. Metab Eng 13:713–722
Zheng Y, Li L, Liu Q, Qin W, Yang J, Cao Y, Jiang X, Zhao G, Xian M (2012) Boosting the free fatty acid synthesis of Escherichia coli by expression of a cytosolic Acinetobacter baylyi thioesterase. Biotechnol Biofuels 5:76
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This work was supported by the National Natural Science Foundation of China (31170040, 31200081) and Chinese Academy of Sciences (KGZD-EW-606).
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Liu, X., Yu, H., Jiang, X. et al. Biosynthesis of butenoic acid through fatty acid biosynthesis pathway in Escherichia coli . Appl Microbiol Biotechnol 99, 1795–1804 (2015). https://doi.org/10.1007/s00253-014-6233-2
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DOI: https://doi.org/10.1007/s00253-014-6233-2