设计一种形式脱酶,用于NADPH再生.
Wei Ma1, Qiang Geng1, Cheng Chen1
1State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Centre for Biomanufacturing, East China University of Science and Technology, Meilong Road 130, Shanghai, 200237, China.
Chembiochem : a European journal of chemical biology
|July 16, 2023
概括
来自Candida dubliniensis的工程形式脱酶 (FDH) 有效地利用尼古丁胺氨酸二核酸盐 (NADPH). 这种增强的酶显示出显著的催化改进,使强大的NADPH再生可用于生物催化.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 合成生物学 合成生物学
- 绿色化学 绿色化学
背景情况:
- 尼古丁胺胺氨基二核酸 (NADH) 和尼古丁胺胺氨基二核酸 (NADPH) 是生物氧化还原反应中的关键 donor.
- 能有效地再生NAD (P) H对于许多酶的生物转化至关重要.
- 现有的形式脱酶 (FDHs) 经常表现出严格的NAD+依赖性和有限的反应性,阻碍NADPH再生.
研究的目的:
- 为了设计一种新型形式脱酶 (FDH),能够有效地利用尼古丁胺氨酸二核酸盐 (NADP+) 进行强大的NADPH再生.
- 提高FDH的催化效率和稳定性,以便在生物催化过程中应用.
主要方法:
- 使用结构导向的理性和半理性设计方法,从Candida dubliniensis (CdFDH) 来设计FDH.
- 组合性突变发生被用来产生具有改进性质的变体.
- 在不对称的氧化/还原转换中酶动力学和性能的表征.
主要成果:
- 与野生类型酶相比,一种组合突变CdFDH-M4的催化效率 (kcat/Km) 增加了75倍.
- CdFDH-M4没有表现出严格的NAD+偏好,有效地利用NADP+.
- 该工程酶在各种不对称的生物催化过程中得到了成功的应用,实现了从135到986的高辅助因子总周转率 (TTN).
结论:
- 经过工程设计的CdFDH-M4代表了NADP+依赖FDHs的显著进步.
- 这种酶为NADPH再生提供了强大而高效的系统,对于各种NADPH依赖的生物催化应用至关重要.
- 这些发现为更可持续,更有效的酶合成途径铺平了道路.
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