Abstract
Biodiesel is produced worldwide as an alternative energy fuel and substitute for petroleum. Biodiesel is often obtained from vegetable oil, but production of biodiesel from plants requires additional land for growing crops and can affect the global food supply. Consequently, it is necessary to develop appropriate microorganisms for the development of an alternative biodiesel feedstock. Escherichia coli is suitable for the production of biodiesel feedstocks since it can synthesize fatty acids for lipid production, grows well, and is amenable to genetic engineering. Recombinant E. coli was designed and constructed for the production of biodiesel with improved unsaturated fatty acid contents via regulation of the FAS pathway consisting of initiation, elongation, and termination steps. Here, we investigated the effects of fabA, fabB, and fabF gene expression on the production of unsaturated fatty acids and observed that the concentration of cis-vaccenic acid, a major component of unsaturated fatty acids, increased 1.77-fold compared to that of the control strain. We also introduced the genes which synthesize malonyl-ACP used during initiation step of fatty acid synthesis and the genes which produce free fatty acids during termination step to study the effect of combination of genes in elongation step and other steps. The total fatty acid content of this strain increased by 35.7% compared to that of the control strain. The amounts of unsaturated fatty acids and cis-vaccenic acid increased by 3.27 and 3.37-fold, respectively.
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Karmakar, A., S. Karmakar, and S. Mukherjee (2010) Properties of various plants and animals feedstocks for biodiesel production. Bioresour. Technol. 101: 7201–7210.
Refaat, A. (2009) Correlation between the chemical structure of biodiesel and its physical properties. Int. J. Environ. Sci. Te. 6: 677–694.
Bringe, N. A., G. Knothe, J. Van Gerpen, and J. Krahl (2005) Soybean Oil Composition For Biodiesel. pp. 161–164. AOCS Press, Champaign, IL, USA.
Kinney, A. and T. Clemente (2005) Modifying soybean oil for enhanced performance in biodiesel blends. Fuel Proc. Technol. 86: 1137–1147.
Knothe, G. (2008) “Designer” biodiesel: Optimizing fatty ester composition to improve fuel properties. Energ. Fuel 22: 1358–1364.
Steen, E. J., Y. Kang, G. Bokinsky, Z. Hu, A. Schirmer, A. McClure, S. B. Del Cardayre, and J. D. Keasling (2010) Microbial production of fatty-acid-derived fuels and chemicals from plant biomass. Nature 463: 559–562.
Magnuson, K., S. Jackowski, C. O. Rock, and J. E. Cronan (1993) Regulation of fatty acid biosynthesis in Escherichia coli. Microbiol. Rev. 57: 522–542.
Cao, Y., J. Yang, M. Xian, X. Xu, and W. Liu (2010) Increasing unsaturated fatty acid contents in Escherichia coli by coexpression of three different genes. Appl. Microbiol. Biot. 87: 271–280.
Meng, X., J. Yang, X. Xu, L. Zhang, Q. Nie, and M. Xian (2009) Biodiesel production from oleaginous microorganisms. Renew. Energ. 34: 1–5.
Handke, P., S. A. Lynch, and R. T. Gill (2011) Application and engineering of fatty acid biosynthesis in Escherichia coli for advanced fuels and chemicals. Metab. Eng. 13: 28–37.
Lu, Y.-J., Y.-M. Zhang, and C. O. Rock (2004) Product diversity and regulation of type II fatty acid synthases. Biochem. Cell Biol. 82: 145–155.
Cho, H. and J. E. Cronan (1995) Defective export of a periplasmic enzyme disrupts regulation of fatty acid synthesis. J. Biol. Chem. 270: 4216–4219.
Teixeira, H., M. Goncalves, N. Rozes, A. Ramos, and M. San Romao (2002) Lactobacillic acid accumulation in the plasma membrane of Oenococcus oeni: A response to ethanol stress? Microb. Ecol. 43: 146–153.
Grogan, D. W. and J. E. Cronan (1997) Cyclopropane ring formation in membrane lipids of bacteria. Microbiol. Mol. Biol. R. 61: 429–441.
Lee, S., E. Jeon, Y. Jung, and J. Lee (2012) Heterologous coexpression of accA, fabD, and thioesterase genes for improving long-chain fatty acid production in Pseudomonas aeruginosa and Escherichia coli. Appl. Biochem. Biotech. 167: 24–38.
Lee, S., S. Park, and J. Lee (2013) Improvement of free fatty acid production in Escherichia coli using codon-optimized Streptococcus pyogenes acyl-ACP thioesterase. Bioproc. Biosyst. Eng. 36: 1519–1525.
Liu, T., H. Vora, and C. Khosla (2010) Quantitative analysis and engineering of fatty acid biosynthesis in E. coli. Metab. Eng. 12: 378–386.
Kyoto Encyclopedia of Genes and Genomes (KEGG). http://www.genome.jp/kegg/.
Sambrook, J. and W. R. David (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.
Lepage, G. and C. C. Roy (1984) Improved recovery of fatty acid through direct transesterification without prior extraction or purification. J. Lipid Res. 25: 1391–1396.
Ramos, M. J., C. M. Fernández, A. Casas, L. Rodríguez, and Á. Pérez (2009) Influence of fatty acid composition of raw materials on biodiesel properties. Bioresour. Technol. 100: 261–268.
Knothe, G. (2010) Biodiesel: Current trends and properties. Top. Catal. 53: 714–720.
Knothe, G. (2005) Dependence of biodiesel fuel properties on the structure of fatty acid alkyl esters. Fuel Proc. Technol. 86: 1059–1070.
Bamgboye, A. and A. Hansen (2008) Prediction of cetane number of biodiesel fuel from the fatty acid methyl ester (FAME) composition. Int. Agrophys. 22: 21.
Pinzi, S., I. Garcia, F. Lopez-Gimenez, M. Luque de Castro, G. Dorado, and M. Dorado (2009) The ideal vegetable oil-based biodiesel composition: A review of social, economical and technical implications. Energ. Fuel 23: 2325–2341.
Garwin, J., A. Klages, and J. E. Cronan (1980) Beta-ketoacylacyl carrier protein synthase II of Escherichia coli. Evidence for function in the thermal regulation of fatty acid synthesis. J. Biol. Chem. 255: 3263–3265.
Marr, A. G. and J. L. Ingraham (1962) Effect of temperature on the composition of fatty acids in Escherichia coli. J. Bacteriol. 84: 1260–1267.
Davis, M. S. and J. E. Cronan (2001) Inhibition of Escherichia coli acetyl coenzyme a carboxylase by acyl-acyl carrier protein. J. Bacteriol. 183: 1499–1503.
Heath, R. J. and C. O. Rock (1996) Regulation of fatty acid elongation and initiation by acyl-acyl carrier protein in Escherichia coli. J. Biol. Chem. 271: 1833–1836.
Davis, M. S., J. Solbiati, and J. E. Cronan (2000) Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J. Biol. Chem. 275: 28593–28598.
Chang, Y.-Y., J. Eichel, and J. E. Cronan (2000) Metabolic instability of Escherichia coli cyclopropane fatty acid synthase is due to RpoH-dependent proteolysis. J. Bacteriol. 182: 4288–4294.
Lu, X., H. Vora, and C. Khosla (2008) Overproduction of free fatty acids in E. coli: implications for biodiesel production. Metab. Eng. 10: 333–339.
Liu, H., C. Yu, D. Feng, T. Cheng, X. Meng, W. Liu, H. Zou, and M. Xian (2012) Production of extracellular fatty acid using engineered Escherichia coli. Microb. Cell Fact. 11: 41.
Meng, X., J. Yang, Y. Cao, L. Li, X. Jiang, X. Xu, W. Liu, M. Xian, and Y. Zhang (2011) Increasing fatty acid production in E. coli by simulating the lipid accumulation of oleaginous microorganisms. J. Ind. Microbiol. Biot. 38: 919–925.
Duan, Y., Z. Zhu, K. Cai, X. Tan, and X. Lu (2011) De novo biosynthesis of biodiesel by Escherichia coli in optimized fedbatch cultivation. Plos One 6: e20265.
Kalscheuer, R., T. Stölting, and A. Steinbüchel (2006) Microdiesel: Escherichia coli engineered for fuel production. Microbiol. 152: 2529–2536.
Zhang, Y. M., H. Marrakchi, and C. O. Rock (2002) The FabR (YijC) transcription factor regulates unsaturated fatty acid biosynthesis in Escherichia coli. J. Biol. Chem. 277: 15558–15565.
Knothe, G., R. O. Dunn, and M. O. Bagby (1997) Biodiesel: The Use Of Vegetable Oils And Their Derivatives As Alternative Diesel Fuels. pp. 172–208. ACS Publications, Washington, DC, USA.
Knothe, G. and R. O. Dunn (2003) Dependence of oil stability index of fatty compounds on their structure and concentration and presence of metals. J. Am. Oil Chem. Soc. 80: 1021–1026.
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Do, K.H., Park, H.M., Kim, S.K. et al. Production of cis-Vaccenic Acid-oriented Unsaturated Fatty Acid in Escherichia coli. Biotechnol Bioproc E 23, 100–107 (2018). https://doi.org/10.1007/s12257-017-0473-9
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DOI: https://doi.org/10.1007/s12257-017-0473-9