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Enhancement of free fatty acid production in Saccharomyces cerevisiae by control of fatty acyl-CoA metabolism.
Appl Microbiol Biotechnol. 2014 Aug; 98(15):6739-50.AM

Abstract

Production of biofuels derived from microbial fatty acids has attracted great attention in recent years owing to their potential to replace petroleum-derived fuels. To be cost competitive with current petroleum fuel, flux toward the direct precursor fatty acids needs to be enhanced to approach high yields. Herein, fatty acyl-CoA metabolism in Saccharomyces cerevisiae was engineered to accumulate more free fatty acids (FFA). For this purpose, firstly, haploid S. cerevisiae double deletion strain △faa1△faa4 was constructed, in which the genes FAA1 and FAA4 encoding two acyl-CoA synthetases were deleted. Then the truncated version of acyl-CoA thioesterase ACOT5 (Acot5s) encoding Mus musculus peroxisomal acyl-CoA thioesterase 5 was expressed in the cytoplasm of the strain △faa1△faa4. The resulting strain △faa1△faa4 [Acot5s] accumulated more extracellular FFA with higher unsaturated fatty acid (UFA) ratio as compared to the wild-type strain and double deletion strain △faa1△faa4. The extracellular total fatty acids (TFA) in the strain △faa1△faa4 [Acot5s] increased to 6.43-fold as compared to the wild-type strain during the stationary phase. UFA accounted for 42 % of TFA in the strain △faa1△faa4 [Acot5s], while no UFA was detected in the wild-type strain. In addition, the expression of Acot5s in △faa1△faa4 restored the growth, which indicates that FFA may not be the reason for growth inhibition in the strain △faa1△faa4. RT-PCR results demonstrated that the de-repression of fatty acid synthesis genes led to the increase of extracellular fatty acids. The study presented here showed that through control of the acyl-CoA metabolism by deleting acyl-CoA synthetase and expressing thioesterase, more FFA could be produced in S. cerevisiae, demonstrating great potential for exploitation in the platform of microbial fatty acid-derived biofuels.

Authors+Show Affiliations

School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, 637459, Singapore, Singapore.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24769906

Citation

Chen, Liwei, et al. "Enhancement of Free Fatty Acid Production in Saccharomyces Cerevisiae By Control of Fatty acyl-CoA Metabolism." Applied Microbiology and Biotechnology, vol. 98, no. 15, 2014, pp. 6739-50.
Chen L, Zhang J, Lee J, et al. Enhancement of free fatty acid production in Saccharomyces cerevisiae by control of fatty acyl-CoA metabolism. Appl Microbiol Biotechnol. 2014;98(15):6739-50.
Chen, L., Zhang, J., Lee, J., & Chen, W. N. (2014). Enhancement of free fatty acid production in Saccharomyces cerevisiae by control of fatty acyl-CoA metabolism. Applied Microbiology and Biotechnology, 98(15), 6739-50. https://doi.org/10.1007/s00253-014-5758-8
Chen L, et al. Enhancement of Free Fatty Acid Production in Saccharomyces Cerevisiae By Control of Fatty acyl-CoA Metabolism. Appl Microbiol Biotechnol. 2014;98(15):6739-50. PubMed PMID: 24769906.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhancement of free fatty acid production in Saccharomyces cerevisiae by control of fatty acyl-CoA metabolism. AU - Chen,Liwei, AU - Zhang,Jianhua, AU - Lee,Jaslyn, AU - Chen,Wei Ning, Y1 - 2014/04/26/ PY - 2014/01/28/received PY - 2014/04/05/accepted PY - 2014/04/04/revised PY - 2014/4/29/entrez PY - 2014/4/29/pubmed PY - 2015/3/10/medline SP - 6739 EP - 50 JF - Applied microbiology and biotechnology JO - Appl Microbiol Biotechnol VL - 98 IS - 15 N2 - Production of biofuels derived from microbial fatty acids has attracted great attention in recent years owing to their potential to replace petroleum-derived fuels. To be cost competitive with current petroleum fuel, flux toward the direct precursor fatty acids needs to be enhanced to approach high yields. Herein, fatty acyl-CoA metabolism in Saccharomyces cerevisiae was engineered to accumulate more free fatty acids (FFA). For this purpose, firstly, haploid S. cerevisiae double deletion strain △faa1△faa4 was constructed, in which the genes FAA1 and FAA4 encoding two acyl-CoA synthetases were deleted. Then the truncated version of acyl-CoA thioesterase ACOT5 (Acot5s) encoding Mus musculus peroxisomal acyl-CoA thioesterase 5 was expressed in the cytoplasm of the strain △faa1△faa4. The resulting strain △faa1△faa4 [Acot5s] accumulated more extracellular FFA with higher unsaturated fatty acid (UFA) ratio as compared to the wild-type strain and double deletion strain △faa1△faa4. The extracellular total fatty acids (TFA) in the strain △faa1△faa4 [Acot5s] increased to 6.43-fold as compared to the wild-type strain during the stationary phase. UFA accounted for 42 % of TFA in the strain △faa1△faa4 [Acot5s], while no UFA was detected in the wild-type strain. In addition, the expression of Acot5s in △faa1△faa4 restored the growth, which indicates that FFA may not be the reason for growth inhibition in the strain △faa1△faa4. RT-PCR results demonstrated that the de-repression of fatty acid synthesis genes led to the increase of extracellular fatty acids. The study presented here showed that through control of the acyl-CoA metabolism by deleting acyl-CoA synthetase and expressing thioesterase, more FFA could be produced in S. cerevisiae, demonstrating great potential for exploitation in the platform of microbial fatty acid-derived biofuels. SN - 1432-0614 UR - https://www.unboundmedicine.com/medline/citation/24769906/Enhancement_of_free_fatty_acid_production_in_Saccharomyces_cerevisiae_by_control_of_fatty_acyl_CoA_metabolism_ L2 - https://dx.doi.org/10.1007/s00253-014-5758-8 DB - PRIME DP - Unbound Medicine ER -