title: Fumaric acid production in Saccharomyces cerevisiae by simultaneous use of oxidative and reductive routes.
abstract: In this study, the simultaneous use of reductive and oxidative routes to produce fumaric acid was explored. The strain FMME003 (Saccharomyces cerevisiae CEN.PK2-1CΔTHI2) exhibited capability to accumulate pyruvate and was used for fumaric acid production. The fum1 mutant FMME004 could produce fumaric acid via oxidative route, but the introduction of reductive route derived from Rhizopus oryzae NRRL 1526 led to lower fumaric acid production. Analysis of the key factors associated with fumaric acid production revealed that pyruvate carboxylase had a low degree of control over the carbon flow to malic acid. The fumaric acid titer was improved dramatically when the heterologous gene RoPYC was overexpressed and 32 μg/L of biotin was added. Furthermore, under the optimal carbon/nitrogen ratio, the engineered strain FMME004-6 could produce up to 5.64 ± 0.16 g/L of fumaric acid. These results demonstrated that the proposed fermentative method is efficient for fumaric acid production.
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China; The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Xu
authors.forename: Xiulai
authors.initials: X
authors.lastname: Chen
authors.forename: Liming
authors.initials: L
authors.lastname: Liu
authors.forename: Linghuo
authors.initials: L
authors.lastname: Jiang
mesh_terms: D001710:Biotin / Q000494:pharmacology; D002244:Carbon / Q000378:metabolism; D005285:Fermentation / Q000187:drug effects; D005650:Fumarates / Q000378:metabolism*; D005818:Genetic Engineering; D008293:Malates / Q000494:pharmacology; D009584:Nitrogen / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D011766:Pyruvate Carboxylase / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D014508:Urea / Q000494:pharmacology
chemical_list: D005650:Fumarates; D008293:Malates; D001710:Biotin; D002244:Carbon; C030298:malic acid; C032005:fumaric acid; D014508:Urea; D011766:Pyruvate Carboxylase; D009584:Nitrogen
keywords: Carbon/nitrogen ratio; Fumaric acid; Oxidative and reductive routes; Pyruvate carboxylase; Saccharomyces cerevisiae