数字农科院2.0

Efficient production of γ-aminobutyric acid using engineered Escherichia coli whole-cell catalyst

文献类型： 外文期刊

作者： Chang F.;Wang Y.;Zhang J.;Tu T.;Luo H.;Huang H.;Bai Y.;Qin X.;Wang Y.;Yao B.;Wang Y.;Wang X.

作者机构：

关键词： Cell permeability;Escherichia coli;PLP self-sufficient system;Whole-cell bioconversion;γ-Aminobutyric acid

期刊名称： Enzyme and Microbial Technology

ISSN： 0141-0229

年卷期： 2024 年 174 卷

页码：

收录情况： SCIE（2024版） ; ; EI（2024版）

摘要： γ-Aminobutyric acid (GABA) has been widely used in the food, feed, pharmaceutical, and chemical industry fields. Previously, we developed a whole-cell catalyst capable of converting L-glutamate (L-Glu) into GABA by overexpressing the glutamate decarboxylase gene (gadz11) from Bacillus sp. Z11 in Escherichia coli BL21(DE3). However, to enhance cell permeability, a freeze-thaw treatment is required, and to enhance GADZ11 activity, pyridoxal 5′-phosphate (PLP) must be added to the reaction system. The aim of this study is to provide a more efficient approach for GABA production by engineering the recombinant E. coli above. First, the inducible expression conditions of the gadz11 in E. coli were optimized to 37 °C for 6 h. Next, an ideal engineered strain was produced via increasing cell permeability by overexpressing sulA and eliminating PLP dependence by constructing a self-sufficient system. Furthermore, an efficient whole-cell biocatalytic process was optimized. The optimal substrate concentration, cell density, and reaction temperature were 1.0 mol/L (the molecular ratio of L-Glu to L-monosodium glutamate (L-MSG) was 4:1), 15 and 37 °C, respectively. Finally, a whole-cell bioconversion procedure was performed in a 3-L bioreactor under optimal conditions. The strain could be reused for at least two cycles with GABA yield, productivity and conversion ratio of 206.2 g/L, 117.8 g/L/h and 100.0%, respectively. This is currently the highest GABA productivity from a mixture of L-Glu and L-MSG reported without the addition of cofactors or additional treatment of cells. This work demonstrates that the novel engineered E. coli strain has the potential for application in large-scale industrial GABA production. © 2023 Elsevier Inc.

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