期刊
BIORESOURCE TECHNOLOGY
卷 148, 期 -, 页码 91-96出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2013.08.115
关键词
Saccharomyces cerevisiae; Fumaric acid; Oxidative and reductive routes; Pyruvate carboxylase; Carbon/nitrogen ratio
资金
- Major State Basic Research Development Program of China [2013CB733600]
- Program for Young Talents in China
- Provincal Outstanding Youth Foundation of Jiangsu Province [BK2012002]
- Fundamental Research Funds for the Central Universities [JUSRP51313B, JUSRP1013]
In this study, the simultaneous use of reductive and oxidative routes to produce fumaric acid was explored. The strain FMME003 (Saccharomyces cerevisiae CEN.PK2-1C Delta THI2) 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 mu g/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. (C) 2013 Elsevier Ltd. All rights reserved.
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