title: Dual regulation of cytoplasmic and mitochondrial acetyl-CoA utilization for improved isoprene production in Saccharomyces cerevisiae.
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.
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Lv
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Wang
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.|Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University, Hangzhou 310027, China.
authors.forename: Lidan
authors.initials: L
authors.lastname: Ye
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Wenping
authors.initials: W
authors.lastname: Xie
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Haoming
authors.initials: H
authors.lastname: Xu
authors.affiliation: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Yu
mesh_terms: D000105:Acetyl Coenzyme A / Q000235:genetics / Q000378:metabolism*; D002070:Butadienes; D003593:Cytoplasm / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D045782:Hemiterpenes / Q000096:biosynthesis*; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways; D008798:Mevalonic Acid / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology*; D010420:Pentanes; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism
chemical_list: D002070:Butadienes; D045782:Hemiterpenes; D010420:Pentanes; C005059:isoprene; D000105:Acetyl Coenzyme A; D008798:Mevalonic Acid