title: Synergy between methylerythritol phosphate pathway and mevalonate pathway for isoprene production in Escherichia coli.
abstract: Isoprene, a key building block of synthetic rubber, is currently produced entirely from petrochemical sources. In this work, we engineered both the methylerythritol phosphate (MEP) pathway and the mevalonate (MVA) pathway for isoprene production in E. coli. The synergy between the MEP pathway and the MVA pathway was demonstrated by the production experiment, in which overexpression of both pathways improved the isoprene yield about 20-fold and 3-fold, respectively, compared to overexpression of the MEP pathway or the MVA pathway alone. The (13)C metabolic flux analysis revealed that simultaneous utilization of the two pathways resulted in a 4.8-fold increase in the MEP pathway flux and a 1.5-fold increase in the MVA pathway flux. The synergy of the dual pathway was further verified by quantifying intracellular flux responses of the MEP pathway and the MVA pathway to fosmidomycin treatment and mevalonate supplementation. Our results strongly suggest that coupling of the complementary reducing equivalent demand and ATP requirement plays an important role in the synergy of the dual pathway. Fed-batch cultivation of the engineered strain overexpressing the dual pathway resulted in production of 24.0g/L isoprene with a yield of 0.267g/g of glucose. The synergy of the MEP pathway and the MVA pathway also successfully increased the lycopene productivity in E. coli, which demonstrates that it can be used to improve the production of a broad range of terpenoids in microorganisms.
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Yang
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China; Shanghai Research and Development Center of Industrial Biotechnology, Shanghai 201201, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China; Shanghai Research and Development Center of Industrial Biotechnology, Shanghai 201201, China.
authors.forename: Bingbing
authors.initials: B
authors.lastname: Sun
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Gao
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China; Shanghai Research and Development Center of Industrial Biotechnology, Shanghai 201201, China; Jiangsu National Synergetic Innovation Center for Advanced Materials, Nanjing, China. Electronic address: syang@sibs.ac.cn.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
mesh_terms: D002070:Butadienes / Q000302:isolation & purification; D066241:Carbon-13 Magnetic Resonance Spectroscopy / Q000379:methods; D003198:Computer Simulation; D004896:Erythritol / Q000031:analogs & derivatives* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D045782:Hemiterpenes / Q000096:biosynthesis* / Q000302:isolation & purification; D060847:Metabolic Engineering / Q000379:methods; D064688:Metabolic Flux Analysis / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000502:physiology*; D008798:Mevalonic Acid / Q000378:metabolism*; D008954:Models, Biological; D010420:Pentanes / Q000302:isolation & purification; D013403:Sugar Phosphates / Q000378:metabolism*
chemical_list: C114232:2-C-methylerythritol 4-phosphate; D002070:Butadienes; D045782:Hemiterpenes; D010420:Pentanes; D013403:Sugar Phosphates; C005059:isoprene; D004896:Erythritol; D008798:Mevalonic Acid
keywords: Metabolic engineering; Methylerythritol phosphate pathway; Mevalonate pathway; Synergy; Terpenoid