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Increasing unsaturated fatty acid contents in Escherichia coli by coexpression of three different genes

  • Applied Genetics and Molecular Biotechnology
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Abstract

Biodiesel is an interesting alternative energy source and is used as substitute for petroleum-based diesel. Microorganisms have been used for biodiesel production due to their significant environmental and economic benefits. However, few researches have investigated the regulation of fatty acid composition of these microbial diesels. Fatty acid biosynthesis in Escherichia coli has provided a paradigm for other bacteria and plants. By overexpressing two genes (fabA and fabB) associated with unsaturated fatty acid (UFA) synthesis in E. coli, we have engineered an efficient producer of UFAs. Saturated fatty acid (SFA) contents decreased from 50.2% (the control strain) to 34.6% (the recombinant strain overexpressing fabA and fabB simultaneously) and the ratio of cis-vaccenate (18:1Δ11), a major UFA in E. coli, reached 51.1% in this recombinant strain. When an Arabidopsis thaliana thioesterase (AtFatA) was coexpressed with these two genes, 0.19 mmol l−1 fatty acids was produced by this E. coli strain after 18-h culture under shake-flask conditions. Free fatty acids made up about 37.5% of total fatty acid concentration in this final engineered strain carrying fabA, fabB, and AtFatA, and the ratio of UFA/SFA reached 2.3:1. This approach offers a means to improve the fatty acid composition of microdiesel and might pave the way for production of biodiesel equivalents using engineered microorganisms in the near future.

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Acknowledgments

This research was sponsored by CAS 100 Talents Program (KGCX2-YW-801). The authors would like to thank Dr. Yun Fa and Cong Zhang for GC analysis and Dr. Wenna Guan and Cong Wang for GC-MS analysis of fatty acid methyl esters.

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Authors and Affiliations

  1. Qingdao Institute of BioEnergy and Bioprocess Technology, Chinese Academy of Sciences, 266101, Qingdao, China

    Yujin Cao, Jianming Yang, Mo Xian, Xin Xu & Wei Liu

Authors
  1. Yujin Cao
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  2. Jianming Yang
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  3. Mo Xian
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  4. Xin Xu
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  5. Wei Liu
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Corresponding author

Correspondence to Mo Xian.

Additional information

Yujin Cao and Jianming Yang contributed equally to this article.

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Fig. S1

Gas chromatograms of fatty acid methyl esters obtained from different E. coli transformants after 3-h induction. A, The control strain harboring pET30a; B, the pET-fabA transformants; C, the pET-fabB transformants; D, the pET-fabAB transformants; E, the pACYC-TE transformants; F, E. coli strain harboring both pET-fabAB and pACYC-TE. 14:0, myristic acid; 16:1 palmitoleic acid; 16:0, palmitic acid; 18:1, cis-vaccenic acid (GIF 350 kb)

High resolution image file (TIFF 399 kb)

Fig. S2

Gas chromatograms of fatty acid methyl esters obtained from different E. coli transformants after 6 h induction. A, The control strain harboring pET30a; B, the pET-fabA transformants; C, the pET-fabB transformants; D, the pET-fabAB transformants; E, the pACYC-TE transformants; F, E. coli strain harboring both pET-fabAB and pACYC-TE. 14:0, myristic acid; 16:1 palmitoleic acid; 16:0, palmitic acid; 17:0c, cis-methylene-9,10-hexadecanoic acid; 18:1, cis-vaccenic acid; 19:0c, cis-methylene-11,12-octadecanoic acid (GIF 369 kb)

High resolution image file (TIFF 237 kb)

Fig. S3

Gas chromatograms of fatty acid methyl esters obtained from different E. coli transformants after 12 h induction. A, The control strain harboring pET30a; B, the pET-fabA transformants; C, the pET-fabB transformants; D, the pET-fabAB transformants; E, the pACYC-TE transformants; F, E. coli strain harboring both pET-fabAB and pACYC-TE. 14:0, myristic acid; 16:1 palmitoleic acid; 16:0, palmitic acid; 17:0c, cis-methylene-9,10-hexadecanoic acid; 18:1, cis-vaccenic acid; 19:0c, cis-methylene-11,12-octadecanoic acid (GIF 399 kb)

High resolution image file (TIFF 425 kb)

Fig. S4

Gas chromatograms of free fatty acids obtained from different E. coli transformants. A, E. coli strain harboring pACYC-TE; B, E. coli strain harboring both pET-fabAB and pACYC-TE. 14:0, myristic acid; 16:1 palmitoleic acid; 16:0, palmitic acid; 18:1, cis-vaccenic acid (GIF 134 kb)

High resolution image file (TIFF 161 kb)

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Cao, Y., Yang, J., Xian, M. et al. Increasing unsaturated fatty acid contents in Escherichia coli by coexpression of three different genes. Appl Microbiol Biotechnol 87, 271–280 (2010). https://doi.org/10.1007/s00253-009-2377-x

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  • DOI: https://doi.org/10.1007/s00253-009-2377-x

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