Journal
JOURNAL OF BIOTECHNOLOGY
Volume 239, Issue -, Pages 90-97Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2016.10.013
Keywords
Branched-chain esters; Ehrlich degradation pathway; Pyruvate carrier; Pyruvate shunt; Pathway segmentation; Saccharomyces cerevisiae
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Funding
- National University of Singapore (NUS Grant) [R279 000 364 133]
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Medium branched-chain esters can be used not only as a biofuel but are also useful chemicals with various industrial applications. The development of economically feasible and environment friendly bio-based fuels requires efficient cell factories capable of producing desired products in high yield. Herein, we sought to use a number of strategies to engineer Saccharomyces cerevisiae for high-level production of branched-chain esters. Mitochondrion-based expression of ATF1 gene in a base strain with an overexpressed valine biosynthetic pathway together with expression of mitochondrion-relocalized alpha-ketoacid decarboxylase (encoded by ARO10) and alcohol dehydrogenase (encoded by ADH7) not only produced isobutyl acetate, but also 3-methyl-1-butyl acetate and 2-methyl-1-butyl acetate. Further segmentation of the downstream esterification step into the cytosol to utilize the cytosolic acetyl-CoA pool for acetyltransferase (ATF)-mediated condensation enabled an additional fold improvement of ester productions. The best titre attained in the present study is 260.2 mg/L isobutyl acetate, 296:1 mg/L 3-methyl-1-butyl acetate and 289.6 mg/L 2-methyl-1-butyl acetate. (C) 2016 Elsevier B.V. All rights reserved.
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