计算机辅助的设计新型纤维素二胺酶,以使用乳糖有效合成乳糖
Dong-Xu Jia1,2, Hai Yu1,2, Fan Wang1,2
1The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, 18 Chaowang Road, Hangzhou, 310014, People's Republic of China.
Bioprocess and biosystems engineering
|July 14, 2023
概括
一种新型的酶 - - 纤维素-2-表皮酶 (CE) - - 为增强乳糖合成而设计. 突变D226G的稳定性和效率有所提高,提高了工业应用的乳糖产量.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酶工程是什么? 酶工程是什么?
- 碳水化合物化学 碳水化合物化学
背景情况:
- 纤维素2-表皮酶 (CE) 对于从乳糖中合成乳糖至关重要.
- 现有的CE受热稳定性和低催化效率的影响,限制了工业用途.
研究的目的:
- 发现和设计一种来自 Caldicellulosiruptor morganii (CmCE) 的新型CE,以改善乳糖的生产.
- 通过向突变发生增强CmCE的稳定性和催化性能.
主要方法:
- 生物信息学发现CmCE.
- 使用FoldX ΔΔG计算和结构分析来创建一个突变库的蛋白质工程.
- 酶动力学和热稳定性的表征.
- 分子建模以阐明突变效应.
主要成果:
- 在α8/α9循环中的突变D226G显著改善了热稳定性 (在65-75°C的半衰期更长).
- D226G表现出较低的Km和较高的kcat/Km值的增强催化效率.
- D226G突变使乳糖合成产量增加到45.5%,而野生型CmCE的产量为35.9%.
结论:
- 设计灵活的α8/α9循环可以增强CE稳定性和催化活性.
- 突变D226G代表了一种有前途的生物催化剂,用于高效的乳糖生物合成.
- 这项研究为改进工业酶应用重新设计CE提供了洞察力.
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