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.
Bioresour Technol. 2013 Nov;148:91-6. doi: 10.1016/j.biortech.2013.08.115. Epub 2013 Aug 26.
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.
在这项研究中，探索了同时使用还原和氧化途径来生产富马酸。菌株 FMME003（酿酒酵母 CEN.PK2-1CΔTHI2）表现出积累丙酮酸的能力，并被用于富马酸生产。 fum1 突变体 FMME004 可以通过氧化途径生产富马酸，但引入来自米根霉 NRRL 1526 的还原途径导致富马酸产量降低。对与富马酸生产相关的关键因素的分析表明，丙酮酸羧化酶对碳流向苹果酸的控制程度较低。当异源基因 RoPYC 过表达并添加 32μg/L 的生物素时，富马酸的产量显著提高。此外，在最佳碳/氮比下，工程菌株 FMME004-6 可以生产高达 5.64±0.16g/L 的富马酸。这些结果表明，所提出的发酵方法可有效用于富马酸生产。
