title: Combinatorial metabolic engineering of Saccharomyces cerevisiae for terminal alkene production.
abstract: Biological production of terminal alkenes has garnered a significant interest due to their industrial applications such as lubricants, detergents and fuels. Here, we engineered the yeast Saccharomyces cerevisiae to produce terminal alkenes via a one-step fatty acid decarboxylation pathway and improved the alkene production using combinatorial engineering strategies. In brief, we first characterized eight fatty acid decarboxylases to enable and enhance alkene production. We then increased the production titer 7-fold by improving the availability of the precursor fatty acids. We additionally increased the titer about 5-fold through genetic cofactor engineering and gene expression tuning in rich medium. Lastly, we further improved the titer 1.8-fold to 3.7 mg/L by optimizing the culturing conditions in bioreactors. This study represents the first report of terminal alkene biosynthesis in S. cerevisiae, and the abovementioned combinatorial engineering approaches collectively increased the titer 67.4-fold. We envision that these approaches could provide insights into devising engineering strategies to improve the production of fatty acid-derived biochemicals in S. cerevisiae.
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 14 Medical Drive, Singapore 117599, Singapore; NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore.
authors.forename: Binbin
authors.initials: B
authors.lastname: Chen
authors.affiliation: NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore; Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117585, Singapore.
authors.forename: Dong-Yup
authors.initials: DY
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 14 Medical Drive, Singapore 117599, Singapore; NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore. Electronic address: bchcmw@nus.edu.sg.
authors.forename: Matthew Wook
authors.initials: MW
authors.lastname: Chang
mesh_terms: D000475:Alkenes / Q000378:metabolism*; D019149:Bioreactors; D005230:Fatty Acids, Nonesterified / Q000096:biosynthesis; D006418:Heme / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D060847:Metabolic Engineering; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
chemical_list: D000475:Alkenes; D005230:Fatty Acids, Nonesterified; D006418:Heme; D006861:Hydrogen Peroxide
keywords: Cofactor; Fatty acid; Gene expression; Metabolic engineering; Saccharomyces cerevisiae; Terminal alkenes