Dual regulation of cytoplasmic and mitochondrial acetyl-CoA utilization for improved isoprene production in Saccharomyces cerevisiae

Xiaomei Lv, Fan Wang, Pingping Zhou, Lidan Ye, Wenping Xie, Haoming Xu and Hongwei Yu ()
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Xiaomei Lv: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University
Fan Wang: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University
Pingping Zhou: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University
Lidan Ye: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University
Wenping Xie: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University
Haoming Xu: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University
Hongwei Yu: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University

Nature Communications, 2016, vol. 7, issue 1, 1-12

Abstract: Abstract Microbial production of isoprene from renewable feedstock is a promising alternative to traditional petroleum-based processes. Currently, efforts to improve isoprenoid production in Saccharomyces cerevisiae mainly focus on cytoplasmic engineering, whereas comprehensive engineering of multiple subcellular compartments is rarely reported. Here, we propose dual metabolic engineering of cytoplasmic and mitochondrial acetyl-CoA utilization to boost isoprene synthesis in S. cerevisiae. This strategy increases isoprene production by 2.1-fold and 1.6-fold relative to the recombinant strains with solely mitochondrial or cytoplasmic engineering, respectively. By combining a modified reiterative recombination system for rapid pathway assembly, a two-phase culture process for dynamic metabolic regulation, and aerobic fed-batch fermentation for sufficient supply of acetyl-coA and carbon, we achieve 2527, mg l−1 of isoprene, which is the highest ever reported in engineered eukaryotes. We propose this strategy as an efficient approach to enhancing isoprene production in yeast, which might open new possibilities for bioproduction of other value-added chemicals.

Date: 2016
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DOI: 10.1038/ncomms12851

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