Enhancing the catalytic efficiency and thermostability of NADH oxidase by a multi-strategy computational approach.

IF 4.2 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

World journal of microbiology & biotechnology Pub Date : 2025-09-26 DOI:10.1007/s11274-025-04553-x

Taisong Shen, Yujiao Li, Hongling Shi, Baomin Luo, Zixing Dong, Yunchao Kan, Dandan Li, Lunguang Yao, Cunduo Tang

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引用次数: 0

Abstract

NADH oxidase (NOX) plays a crucial role in diverse fields, including medicine, food processing, and environmental protection, by catalyzing the oxidation of NADH to NAD⁺. This reaction is essential for biosensor performance, metabolic regulation, anti-aging processes, and the degradation of industrial pollutants. In this study, a NOX (EfNOX) was obtained through gene cloning from Enterococcus faecium. However, its limited thermal stability and catalytic efficiency constrained its industrial applications. To address these limitations, rational design strategies were employed to generate a double mutant, N211M/Q293L. The mutant exhibited a half-life of 27 min at 50℃ and a specific activity of 24.9 U/mg, representing 2.08-fold and 1.87-fold improvements, respectively, over the wild-type EfNOX. These enhancements are expected to promote the applicability of EfNOX in high-temperature environments and its industrial application potential. Collectively, this work offers a practical and efficient strategy for enhancing NOX and other industrial enzymes.

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用多策略计算方法提高NADH氧化酶的催化效率和热稳定性。

NADH氧化酶（NOX）通过催化NADH氧化生成NAD⁺，在医药、食品加工、环境保护等多个领域发挥着至关重要的作用。这种反应对于生物传感器性能、代谢调节、抗衰老过程和工业污染物的降解至关重要。本研究通过对粪肠球菌的基因克隆获得了一种NOX （EfNOX）。然而，其热稳定性和催化效率有限，限制了其工业应用。为了解决这些限制，采用合理的设计策略产生了双突变体N211M/Q293L。该突变体在50℃下的半衰期为27 min，比活性为24.9 U/mg，分别是野生型EfNOX的2.08倍和1.87倍。这些改进有望提高EfNOX在高温环境中的适用性及其工业应用潜力。总的来说，这项工作为增强氮氧化物和其他工业酶提供了一种实用而有效的策略。

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

World journal of microbiology & biotechnology 工程技术-生物工程与应用微生物

CiteScore

6.30

自引率

2.40%

发文量

257

审稿时长

2.5 months

期刊介绍： World Journal of Microbiology and Biotechnology publishes research papers and review articles on all aspects of Microbiology and Microbial Biotechnology. Since its foundation, the Journal has provided a forum for research work directed toward finding microbiological and biotechnological solutions to global problems. As many of these problems, including crop productivity, public health and waste management, have major impacts in the developing world, the Journal especially reports on advances for and from developing regions. Some topics are not within the scope of the Journal. Please do not submit your manuscript if it falls into one of the following categories: · Virology · Simple isolation of microbes from local sources · Simple descriptions of an environment or reports on a procedure · Veterinary, agricultural and clinical topics in which the main focus is not on a microorganism · Data reporting on host response to microbes · Optimization of a procedure · Description of the biological effects of not fully identified compounds or undefined extracts of natural origin · Data on not fully purified enzymes or procedures in which they are applied All articles published in the Journal are independently refereed.