通过循环德克斯介导稳定和prenyltransferase工程进行的 cannabigerolic 酸的综合级联生物催化
Ye Chan Kim1, Hyunseok Yoon1, Youngwoo Na1
1Department of Systems Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul-050-29, South Korea.
Bioresource technology
|August 27, 2025
概括
通过综合生物工艺实现了可持续生产的 cannabigerolic 酸 (CBGA). 这种方法提高了可扩展的大麻素前体生物合成的酶稳定性和辅助因子效率.
科学领域:
- 生物技术
- 合成生物学
- 生物催化
背景情况:
- 大麻酸 (CBGA) 是治疗性大麻素的重要前体.
- 由于酶的不稳定性和高的辅助因子成本,目前的生产方法面临限制.
研究的目的:
- 为CBGA生物合成开发一种高效和可持续的生物过程.
- 克服工业规模生产的酶稳定性和辅助因子依赖性的挑战.
主要方法:
- 综合生物工艺设计,包括循环胺介导稳定.
- 使用基生物传感器来识别和解决基酸盐供应的速度限制步骤.
- 采用NphB酶的组合工程,并结合了腺三酸盐 (ATP) 再生系统.
- 通过体外和体内 (大肠杆菌) 生产验证了该过程.
主要成果:
- 通过循环氨酸稳定实现了体外CBGA产量的7倍增加 (至4mM).
- 改造后的NphB变种显示出明显改善的动力参数 (Km低8.8倍,kcat高1.5倍).
- 通过再生系统证明了5mM的体外CBGA产生与ATP需求的减少.
- 在大肠杆菌中实现了16μMCBGA的微生物生产概念验证.
结论:
- 综合生物工艺为大麻素前体生物合成提供了可扩展和成本效益的平台.
- 结合稳定,途径优化,酶工程和辅助因子管理是可持续工业大麻素生产的关键.
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