A Study on the Efficient Preparation of α-Ketoglutarate with L-Glutamate Oxidase

文献类型： 外文期刊

第一作者： Niu, Shuhui

作者： Niu, Shuhui;Liu, Fang;Rao, Ben;Wang, Yueying;Niu, Shuhui;Wang, Yaping

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关键词： alpha-ketoglutaric acid (alpha-KG); L-glutamate oxidase; catalase; whole-cell catalysis

期刊名称：MOLECULES （ 影响因子：4.6； 五年影响因子：4.9 ）

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年卷期： 2024 年 29 卷 8 期

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收录情况： SCI

摘要： Alpha-ketoglutaric acid (alpha-KG), as an intermediate product of the tricarboxylic acid cycle, plays a crucial role in peptide and amino acid synthesis. In order to reduce costs and improve efficiency in the oxidative production of alpha-ketoglutaric acid, this study successfully synthesized and expressed L-glutamate oxidase (LGOXStr) from Streptomyces viridosporus R111 and catalase (KatGEsc) from Escherichia coli H736. Two immobilization methods and the conditions for one-step whole-cell catalysis of alpha-ketoglutaric acid were investigated. alpha-Ketoglutaric acid has broad applications in the pharmaceutical, food, and chemical industries. The specific research results are as follows: (1) By fusing the sfGFP tag, L-glutamate oxidase (LGOXStr r) and catalase (KatGEsc) were successfully anchored to the outer membrane of Escherichia coli cells, achieving one-step whole-cell catalysis of alpha-ketoglutaric acid with a conversion efficiency of up to 75%. (2) Through the co-immobilization of LGOXStr and KatGEsc, optimization of the preparation parameters of immobilized cells, and exploration of the immobilization method using E.coli@ZIF-8, immobilized cells with conversion rates of over 60% were obtained even after 10 cycles of reuse. Under the optimal conditions, the production rate of alpha-ketoglutaric acid reached 96.7% in a 12 h reaction, which is 1.1 times that of E. coli@SA and 1.29 times that of free cells.

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