对于NADPH再生的高度稳定和可溶性酒精脱酶的计算设计
Jinling Xu1, Haisheng Zhou2, Haoran Yu1,3
1Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, 310027, China.
Bioresources and bioprocessing
|April 23, 2024
概括
工程性酒精脱酶 (ADH) 有效地再生尼古丁胺胺氨基二核酸 (NADPH) 辅因子,促进绿色化学合成. 这种增强的酶在工业应用中表现出更好的活性和稳定性.
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?
- 绿色化学 绿色化学
背景情况:
- 尼古丁胺胺氨基二核酸 (NADPH) 对于工业催化中的酶性氧化还原反应至关重要.
- 能有效地将NADP+再生为NADPH,对于可持续的化学合成至关重要,但仍然是一个挑战.
- 酒精脱酶 (ADH) 酶因其高活性和简单的副产品分离而具有在位辅助因子再生的潜力.
研究的目的:
- 为了改造来自Clostridium beijerinckii的NADP+特异性酒精脱酶 (ADH),以改善辅因子再生.
- 为了提高工业生物催化剂的酶活性,稳定性和整体性能.
主要方法:
- 蛋白质工程使用自动化算法蛋白质修复一站式商店 (PROSS).
- 在复合大肠杆菌中工程酶的定向进化和高细胞密度发酵.
- 酶活性和稳定性测试,包括热稳定性测量.
- 工程酶的应用在合金芳香醇的合成中.
主要成果:
- 与野生类型 (2.9 U/mL) 相比,工程突变CbADH-6M的活性增加了16倍 (46.3 U/mL).
- 观察到增强的热稳定性, ΔT50 的 +3.6 °C.
- 高细胞密度发酵进一步提高了酶活性,达到2401.8U/mL.
- 这种突变酶在NADPH再生中表现出了卓越的效率,用于性酒精合成.
结论:
- 设计的CbADH-6M酶显著提高了NADP+到NADPH再生效率.
- 增强的酶具有高活性和稳定性,显示了绿色化学合成的可观工业潜力.
- 这项工作为可持续生产精细化学品提供了强大的生物催化剂.
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