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结构指导工程对双功能酶NadR进行增强的尼古丁胺氨酸二核酸生产

Dianju Wei¹, Yinfeng Huang¹, Daifu Shi¹

¹Key Laboratory of Bioorganic Synthesis of Zhejiang Province, The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, China.

Biotechnology and bioengineering

|November 5, 2025

概括

研究人员设计了一种单一的酶,可以直接合成尼古丁胺胺氨基二核酸 (NAD+),克服了以前方法的局限性. 这种新的方法提高了用于工业和治疗用途的NAD+生产效率.

科学领域:

生物催化剂是一种生物催化剂.
酵素工程是什么意思酵素工程
代谢工程是代谢工程.

背景情况:

尼古丁胺胺氨基二核酸 (NAD+) 是许多生物过程和工业应用中的重要辅助因子.
目前对NAD+的化学合成方法是危险和低效的.
使用双酶级联的现有酶方法面临着由于分离的活性位点而导致的低催化效率和差的中间道化挑战.

研究的目的:

开发一种直接的,单步生物合成路径,用于NAD+生产.
在双功能酶中增强尼古丁胺基酸酶 (NRK) 域的催化效率.
调查分子内基质通道,以提高生物催化剂性能.

主要方法:

使用Haemophilus influenzae NadR,一种具有NRK和NMNAT活动的自然双功能酶.
开发了一个循环工程对接组合模拟 (LEDCS) 策略,以重新设计NRK领域.
进行结构和相互作用分析以了解酶机制和基质结合.

主要成果:

使用工程双功能酶创建了一个单步NAD+生物合成途径.
V241S/L387Q突变体的酶活性增加了四倍,催化效率提高了2.4倍.
确定了P环在基质结合中的关键作用,并增强了口袋水友性,以改善基质亲和力.

结论:

证明了分子内基质道在多功能酶中的催化优势.
提出了一项可推广的战略,以提高工程生物催化剂的性能.
工程酶为NAD+生物合成提供了更有效和更直接的途径.

关键词:

这是一种 NadR 蛋白质.生物催化剂的生物催化剂分子动力学模拟模拟尼古丁胺腺因二核酸是什么尼古丁胺胺的 рибо酸激酶.在 p-loop 工程方面,

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