精细调整的CRISPRi模块用于升级Bacillus subtilis蛋白质表达系统中的构成性促进体的性能
Tao Liu1, Bin Ye2, Yilan Zhang1
1Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, People's Republic of China.
Journal of agricultural and food chemistry
|December 25, 2025
概括
研究人员开发了一种可酶诱导的CRISPRi系统,以平衡Bacillus subtilis的生长和蛋白质生产. 这种动态控制方法提高了38%的蛋白质产量,提供了改进的工业菌株策略.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 合成生物学 合成生物学
背景情况:
- 细菌是工业蛋白质生产的关键宿主.
- 强大的构成性促进体虽然有效,但会给宿主带来负担,减少生长和生物质.
- 现有的方法缺乏在发酵过程中对促进剂活性的动态控制.
研究的目的:
- 开发一种动态的调节系统,用于控制Bacillus subtilis. 的基因表达.
- 为了克服工业蛋白质生产中强大的构成性促进剂的局限性.
- 为了改善宿主细胞生长和重组蛋白质产量之间的平衡.
主要方法:
- 设计了一个可诱导西洛斯的CRISPR干扰 (CRISPRi) 模块.
- 通过促进器工程和树脂度调整了CRISPRi模块的强度.
- 在发酵过程中对目标促进剂活性实施动态控制.
主要成果:
- 克西洛斯诱导的CRISPRi模块在克西洛斯的存在下成功抑制了向促进物,促进了早期细胞生长.
- 克西洛斯的消费导致促进体强度的恢复,促进蛋白质的表达.
- 与构成性促进体系统相比,目标蛋白产量增加了38%.
- 在细胞生长和蛋白质生产之间表现出更好的平衡.
结论:
- 开发的可酶诱导的CRISPRi系统提供了一种简单有效的方法,可以动态控制Bacillus subtilis的基因表达.
- 这种方法可以通过优化宿主生长和蛋白质生产之间的权衡来提高工业菌株性能.
- 该技术为升级依赖强大的构成性促进剂的工业菌株提供了有价值的工具.
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