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

文献类型： 外文期刊

第一作者： Chang, Fangfang

作者： Chang, Fangfang;Wang, Yuheng;Zhang, Jie;Tu, Tao;Luo, Huiying;Huang, Huoqing;Bai, Yingguo;Qin, Xing;Wang, Yaru;Yao, Bin;Wang, Yuan;Wang, Xiaolu

作者机构：

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

期刊名称：ENZYME AND MICROBIAL TECHNOLOGY （ 影响因子：3.4； 五年影响因子：3.3 ）

ISSN： 0141-0229

年卷期： 2024 年 174 卷

页码：

收录情况： SCI

摘要： gamma-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 degree celsius 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 LGlu to L-monosodium glutamate (L-MSG) was 4:1), 15 and 37 degree celsius, 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.

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