title: Whole-cell biocatalytic and de novo production of alkanes from free fatty acids in Saccharomyces cerevisiae.
abstract: Rapid global industrialization in the past decades has led to extensive utilization of fossil fuels, which resulted in pressing environmental problems due to excessive carbon emission. This prompted increasing interest in developing advanced biofuels with higher energy density to substitute fossil fuels and bio-alkane has gained attention as an ideal drop-in fuel candidate. Production of alkanes in bacteria has been widely studied but studies on the utilization of the robust yeast host, Saccharomyces cerevisiae, for alkane biosynthesis have been lacking. In this proof-of-principle study, we present the unprecedented engineering of S. cerevisiae for conversion of free fatty acids to alkanes. A fatty acid α-dioxygenase from Oryza sativa (rice) was expressed in S. cerevisiae to transform C12-18free fatty acids to C11-17aldehydes. Co-expression of a cyanobacterial aldehyde deformylating oxygenase converted the aldehydes to the desired alkanes. We demonstrated the versatility of the pathway by performing whole-cell biocatalytic conversion of exogenous free fatty acid feedstocks into alkanes as well as introducing the pathway into a free fatty acid overproducer for de novo production of alkanes from simple sugar. The results from this work are anticipated to advance the development of yeast hosts for alkane production. Biotechnol. Bioeng. 2017;114: 232-237. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, Singapore.
authors.forename: Jee Loon
authors.initials: JL
authors.lastname: Foo
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, Singapore.
authors.forename: Adelia Vicanatalita
authors.initials: AV
authors.lastname: Susanto
authors.affiliation: Joint BioEnergy Institute, Emeryville, California.|Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California.|Department of Chemical and Biomolecular Engineering and Department of Bioengineering, University of California, Berkeley, California.
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, Singapore.|Singapore Institute of Technology, Singapore.
authors.forename: Susanna Su Jan
authors.initials: SS
authors.lastname: Leong
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, Singapore.
authors.forename: Matthew Wook
authors.initials: MW
authors.lastname: Chang
mesh_terms: D000473:Alkanes / Q000378:metabolism*; D055162:Biocatalysis; D056804:Biofuels; D019149:Bioreactors / Q000382:microbiology; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism; D005230:Fatty Acids, Nonesterified / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D012275:Oryza / Q000201:enzymology / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
chemical_list: D000473:Alkanes; D056804:Biofuels; D005230:Fatty Acids, Nonesterified; D010940:Plant Proteins; D011994:Recombinant Proteins; D049308:Dioxygenases
keywords: aldehyde; alkane; biofuels; de novo biosynthesis; fatty acid; metabolic engineering; whole-cell biocatalysis