title: Engineering Escherichia coli for fumaric acid production from glycerol.
abstract: The evolved mutant Escherichia coli E2 previously developed for succinate production from glycerol was engineered in this study for fumaric acid production under aerobic conditions. Through deletion of three fumarases, 3.65g/L fumaric acid was produced with the yield of 0.25mol/mol glycerol and a large amount of acetate was accumulated as the main byproduct. In order to reduce acetate production several strategies were attempted, among which increasing the flux of the anaplerotic pathways through overexpression of phosphoenolpyruvate carboxylase gene ppc or the glyoxylate shunt operon aceBA effectively reduced acetate and improved fumaric acid production. In fed-batch culture, the resulting strain EF02(pSCppc) produced 41.5g/L fumaric acid from glycerol with 70% of the maximum theoretical yield and an overall productivity of 0.51g/L/h.
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Zhiwen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan 430068, PR China.
authors.forename: Ya-Jie
authors.initials: YJ
authors.lastname: Tang
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China. Electronic address: chentao@tju.edu.cn.
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Xueming
authors.initials: X
authors.lastname: Zhao
mesh_terms: D000085:Acetates / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology; D061249:Batch Cell Culture Techniques; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D005649:Fumarate Hydratase / Q000378:metabolism; D005650:Fumarates / Q000378:metabolism*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005990:Glycerol / Q000378:metabolism*; D007522:Isocitrate Lyase / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D009097:Multienzyme Complexes / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D010730:Phosphoenolpyruvate Carboxylase / Q000378:metabolism
chemical_list: D000085:Acetates; D005650:Fumarates; D009097:Multienzyme Complexes; C032005:fumaric acid; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D010730:Phosphoenolpyruvate Carboxylase; D007522:Isocitrate Lyase; D005649:Fumarate Hydratase; D005990:Glycerol
keywords: Acetate accumulation; Escherichia coli; Fed-batch culture; Fumaric acid; Glycerol