Alleviating Substrate Inhibition of Leucine Dehydrogenase by Enhancing NAD+ Dissociation Efficiency

IF 3.2 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Biotechnology Journal Pub Date : 2025-06-09 DOI:10.1002/biot.70045

Jun-Ping Zhou, Yi-Nan Xue, Feng Wang, Hui Gao, Zhi-Cheng Zhang, Ai-Ping Pang, Zhi-Qiang Liu, Yu-Guo Zheng

引用次数: 0

Abstract

Leucine dehydrogenase catalyzes the asymmetric reductive amination of ketoacids to produce valuable chiral unnatural amino acids, but its application is often constrained by substrate inhibition. Here we employed a rational engineering strategy to alleviate substrate inhibition by enhancing the dissociation efficiency of NAD⁺. Notably, the variant H184A exhibited a 1.53-fold increase in enzyme activity and a 58% improvement in the half-maximal inhibitory concentration for 2-oxobutyric acid, alleviating substrate inhibition.

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通过提高NAD+解离效率减轻底物对亮氨酸脱氢酶的抑制作用

亮氨酸脱氢酶催化酮酸的不对称还原胺化反应生成有价值的手性非天然氨基酸，但其应用常常受到底物抑制的限制。本文采用合理的工程策略，通过提高NAD+的解离效率来减轻底物的抑制作用。值得注意的是，变异H184A的酶活性增加了1.53倍，对2-氧丁酸的半最大抑制浓度提高了58%，减轻了底物的抑制作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine

CiteScore

8.90

自引率

2.10%

发文量

123

审稿时长

1.5 months

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