设计基于生物传感器的高通量选平台,用于在大肠杆菌中生产高产咖啡酸
Daoguang Tian1, Zhen Qin2, Weilin Liu1
1School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Key Laboratory of Systems Bioengineering, Ministry of Education (Tianjin University), Tianjin, 300072, China; Zhejiang Institute of Tianjin University, Shaoxing, 312300, China.
Metabolic engineering
|October 1, 2025
概括
我们开发了一个由生物传感器驱动的平台,以有效地在大肠杆菌中产生咖啡酸 (CA). 这种方法克服了生产挑战,实现了迄今为止报告的最高CA标位.
科学领域:
- 微生物生物技术 微生物生物技术
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
背景情况:
- 咖啡酸 (CA) 是一种有价值的化合物,具有多种应用.
- 微生物对CA的产生受到低酶活性和产品毒性的阻碍.
研究的目的:
- 为咖啡酸 (CA) 生产开发一个高效的高通量选 (HTS) 平台.
- 克服CA生物合成的瓶,包括酶活性和产品细胞毒性.
主要方法:
- 使用CarR转录因子设计了一种新的p-酸 (p-CA) 生物传感器.
- 将生物传感器与光激活细胞分类 (FACS) 集成为HTS.
- 对大肠杆菌菌株进行了代谢工程,以提高CA的产生.
主要成果:
- 优化了p-CA生物传感器,提高了灵敏度和动态范围.
- 鉴定了一种改进的FjTALG85S突变,具有6.85倍高的催化活性.
- 在5L生物反应器中使用工程菌株CA8.8实现了9.61g/L的CA标位记录.
结论:
- 开发的生物传感器-HTS平台加速了微生物细胞工厂的工程.
- 这项工作为有效生产p-CA和其他衍生化学品提供了强大的工具.
- 成功解决了CA生物合成的关键挑战,例如酶效率和细胞毒性.
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