Expanding the cofactor range of aldehyde reductase YqhD by mutating key amino acids.

IF 2.5 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2026-05-25 DOI:10.1007/s10529-026-03747-z

Xiaohui Wu, Yinghan Hu, Yanzhe Huang, Lingyun Zhang, Yan Xue, Hailong Cao, Xueying Wang, Yongjin J Zhou

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引用次数: 0

Abstract

YqhD is a broad-substrate aldehyde reductase that natively prefers NADPH and is widely used in alcohol biosynthesis. However, insufficient NADPH supply and low cofactor regeneration efficiency limit its catalytic performance. To address this limitation, we engineered YqhD through structure-guided semi-rational design and obtained a mutant G37D with markedly improved dual-cofactor compatibility with NADPH and NADH. With butanal and 3-hydroxypropionaldehyde as substrates, G37D exhibited 2.6- and 4.8-fold increases in catalytic efficiency (k_cat/K_m) with NADH as the cofactor, while retaining > 70% of the wild-type efficiency with NADPH, resulting in comparable efficiencies with both cofactors. Further structural analysis and molecular dynamics simulations revealed that the G37D mutation enhances dual-cofactor utilization by reprogramming the hydrogen-bond network and stabilizing the catalytically favorable closed conformation. This study provides an improved enzyme module for the biomanufacturing of 1,3-propanediol and related alcohols, and offers new insights into the design of versatile catalysts for diverse biocatalytic scenarios.

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通过突变关键氨基酸扩大醛还原酶YqhD的辅因子范围。

YqhD是一种天然倾向于NADPH的宽底物醛还原酶，广泛应用于酒精生物合成。然而，NADPH供应不足和辅助因子再生效率低限制了其催化性能。为了解决这一限制，我们通过结构引导的半理性设计对YqhD进行了改造，并获得了与NADPH和NADH双辅因子相容性显著改善的突变体G37D。以丁醛和3-羟丙醛为底物，G37D以NADH为辅助因子时的催化效率（kcat/Km）分别提高2.6倍和4.8倍，而以NADPH为辅助因子时的催化效率仍保持野生型效率的70%，与这两种辅助因子的效率相当。进一步的结构分析和分子动力学模拟表明，G37D突变通过重编程氢键网络和稳定催化有利的封闭构象，提高了双辅助因子的利用。本研究为1,3-丙二醇及其相关醇的生物合成提供了一种改进的酶模块，并为多种生物催化场景的多用途催化剂的设计提供了新的见解。

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 Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.