在Bacillus subtilis中高效的蛋白质表达和生物合成基因集群调节,由T7-BOOST系统驱动
Yaokang Wu1,2, Yang Li1,2, Yuting Zhang1,2
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.
ACS synthetic biology
|October 27, 2023
概括
研究人员开发了一种T7-BOOST系统,用于精确调节Bacillus subtilis中的基因. 该系统可实现高效的蛋白质和化学生产,为合成生物学应用提供了强大的工具.
科学领域:
- 合成生物学 合成生物学
- 微生物生物技术 微生物生物技术
- 基因规则 基因规则
背景情况:
- 细菌细菌是一种安全的微生物,广泛用于工业.
- 与大肠杆菌相比,B. subtilis中现有的遗传工具是有限的.
- 在B. subtilis中需要先进的遗传调节系统,以实现增强的应用.
研究的目的:
- 在B. subtilis.中构建一个新的,基于T7的,用于转录 (T7-BOOST) 系统的优化输出策略.
- 为了实现低泄漏表达和广泛的基因调节的动态范围.
- 证明该系统在各种应用中的实用性,包括生物催化和蛋白质表达.
主要方法:
- 开发了一个由两个模块组成的T7-BOOST系统,使用可诱导的促进剂P$_{}$和P$_{}$.
- 使用CRISPR/Cpf1系统将T7RNA聚合酶模块集成到B. subtilis基因组中.
- 介绍了表达控制模块在不同副本数 (低,中,高) 的等离子体上进行表征.
主要成果:
- 成功构建并表现出T7-BOOST系统的低泄漏率和广泛的动态范围.
- 证明了全细胞催化为γ-氨基黄油酸的产生 (109.8 g/L,98.0%的转化).
- 成功表达了人体素和人体乳酸,并调节了利科和利博弗拉生物合成基因群.
结论:
- 在B. subtilis. 中,T7-BOOST系统提供了严格和可控制的基因表达.
- 该系统是微生物生物技术中各种应用的有效工具.
- 插即用性质增强了它对合成生物学的多功能性.
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