通过合理设计,增强乳酸脱酶的催化活性和稳定性,用于D-PLA的级联催化
Yufeng Zhou1, Xiaolong Sun2, Jiahuan Hu2
1Taizhou Key Laboratory of Biomass Functional Materials Development and Application, Taizhou University, Jiaojiang, Zhejiang 318000, China; School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, Jiangsu 210023, China.
Journal of biotechnology
|January 7, 2024
概括
研究人员设计了一种突变的乳酸脱酶 (LrLDH-M1) 酶,显著提高了其D-酸生产的效率和稳定性. 这种增强的酶使新的合催化系统能够改善生物制造.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 生物催化剂是一种生物催化剂.
背景情况:
- D-乳酸 (D-PLA) 是一种有价值的中间体,在医学和环保防腐剂中具有应用.
- 有效和成本效益的生产方法对于D-PLA的工业可行性至关重要.
- 乳酸脱酶 (LDH) 在D-PLA合成中起着关键作用,但本地酶往往缺乏最佳的催化效率和稳定性.
研究的目的:
- 合理设计和工程乳酸脱酶 (LDH) 以提高催化效率和稳定性.
- 为了研究工程LDH突变的酶性质和结构基础.
- 开发一个强大的合催化系统,以高效地生产D-酸.
主要方法:
- 合理设计和乳酸脱酶 (LDH) 的位点定向突变发生,以创建突变变体.
- 酶测试以确定野生类型和突变酶的运动参数 (Kcat,Km) 和特定活性.
- 热稳定性分析和对恶劣条件 (如酸性环境) 的耐受性评估.
- 分子对接模拟以阐明改善酶动力学的结构基础.
- 使用工程化大肠杆菌菌株用于D-PLA合成的两种细菌合催化系统的开发.
主要成果:
- 一种四重突变酶,LrLDH-M1 (T247I/D249A/F306W/A214Y),在Kcat.cat的特定活性增加了3.3倍,改善了2.08倍.
- 与野生类型相比,突变酶表现出较低的Km和更高的催化效率 (Kcat/Km).
- 在酸性环境中,LrLDH-M1表现出增强的热稳定性和优越的耐受性.
- 一个由两种细菌结合的系统在两步级联反应中实现了L-phenylalanine到D-phenyllactic acid的77.7%的转化率.
结论:
- 工程LrLDH-M1酶具有显著提高的催化效率,稳定性和对恶劣条件的耐受性.
- 分子对接提供了对突变物增强的酶性质的见解.
- 开发的合催化系统为工业生产D-酸提供了一个有希望和高效的方法.
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