作为辅助因子增强系统的西洛斯减少酶在体内生物催化剂的使用
Chalermroj Sutthaphirom1, Pimchai Chaiyen1
1School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
Methods in enzymology
|April 27, 2025
概括
添加西洛斯减少酶和乳糖可增强工程细胞中的辅因子合成. 这种方法提高了生物转化效率和产品产量,为全细胞生物催化剂提供了一种多功能工具.
科学领域:
- 代谢工程是代谢工程.
- 生物催化剂是一种生物催化剂.
- 合成生物学 合成生物学
背景情况:
- 同因子失衡限制了整个细胞的生物转化效率.
- 提高细胞内辅助因子水平 (NAD,P,H,FMN,FAD,ATP,CoA) 的策略至关重要.
- 现有的方法往往涉及复杂的路径工程.
研究的目的:
- 提出一种新型和通用的XR/乳糖系统,以提高辅因子的可用性.
- 为了证明该系统在提高生物转化产量的实用性.
- 为其在工程微生物中的应用提供一个实际的协议.
主要方法:
- 代谢工程的埃舍里希亚大肠杆菌BL21(DE3) 用于脂肪酒精生物合成.
- 在培养基中添加氧化糖还原酶 (XR) 和乳糖.
- 测量细胞内辅助因子水平和产品产量.
主要成果:
- 在XR/乳糖系统显著增加了糖酸盐的细胞内水平.
- 增强的糖酸盐池导致NAD,P) H,FMN,FAD,ATP和CoA的合成增加.
- 这导致生物转化产品的产量更高,特别是脂肪醇.
结论:
- XR/乳糖系统是改善全细胞生物催化剂辅因子供应的有效和可适应的工具.
- 这种方法可以缓解各种工程微生物系统中的辅因子限制.
- 提供的协议有助于实现这种用于增强生物生产的系统.
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