再组蛋白质的高效自诱导表达通过DegSU定数感应系统在一个强大的细菌中
Hong Zong1,2, Liya Zhang1,2, Yiwen Cheng1,2
1School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China.
ACS synthetic biology
|January 6, 2025
概括
研究人员优化了Bacillus subtilis DegSU的质量检测系统,以提高重组蛋白质的生产. 促进器和调节电路的修改显著增加了蛋白质产量和工业价值化合物生产.
科学领域:
- 微生物学 微生物学
- 合成生物学 合成生物学
- 生物技术是生物技术.
背景情况:
- 在 Bacillus subtilis 中的 DegSU 定数感应 (QS) 系统促进了重组蛋白的自我诱导表达.
- 限制包括不够的促进者力量和复杂的监管网络,阻碍了实际应用.
研究的目的:
- 提高DegSU QS系统的效率和强度,以改善Bacillus subtilis的重组蛋白质生产.
- 通过遗传和监管修改,克服本土系统的局限性.
主要方法:
- 启动器工程:通过改变其核心区域,截断上游序列,并添加激活绑定站点来创建QS响应的启动器P进行修改,以创建P_E742.
- 数学建模:开发了一个模型来量化QS系统的监管动态.
- 系统优化:淘汰了竞争基因 (sacB,amyE) 和操作子 (pgs,srfA),在一个强大的B. subtilis菌株中引入了特定的DegU和DegQ变体 (degU^L113F,degQ^36^Hy).
主要成果:
- 与原始促进剂相比,P_E742的促进剂强度增加了118.3%.
- 优化的DegSU QS系统显示,监管强度增加了84.0%.
- 获得了52.5%的氨酸标位增加和65.9%的细胞外氧酶活性增加.
结论:
- 改造的P_E742促进体显著增强了基因表达.
- 以数学建模为指导的DegSU QS系统的优化,大大改善了Bacillus subtilis中有价值的化合物和重组蛋白的产生.
- 这项研究验证了修改后的DegSU QS系统在高水平蛋白质和代谢物生产中的工业潜力.
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