在活体中对Bacillus subtilisBacillus subtilisBacillus subtilisBacillus subtilisBacillus subtilisBacillus subtilisBacillus
Maliheh Vahidinasab1, Lisa Thewes1, Bahar Abrishamchi1
1Department of Bioprocess Engineering (150k), Institute of Food Science and Biotechnology, University of Hohenheim, Fruwirthstrasse 12, 70599 Stuttgart, Germany.
Microorganisms
|November 27, 2024
概括
这项研究开发了一种新的体内系统,用于量化Bacillus subtilis中形成表面因子的非核糖体合成酶 (NRPS) 的可用性. 这些发现揭示了生物过程中的NRPS动态,有助于优化生物表面活性剂的生产.
科学领域:
- 生物技术是生物技术.
- 微生物生理学 微生物生理学
- 合成生物学 合成生物学
背景情况:
- 表面活性素是Bacillus subtilis的一个关键生物表面活性剂,由由srfAA-AD操作子编码的非核糖体合成酶 (NRPS) 复合物合成.
- 在生物过程中这种NRPS复合物的体内动力学和量化程度尚不清楚,尽管它与定数感应有关.
研究的目的:
- 建立一个体内定量系统,用于监测Bacillus subtilis中单个表面素形成NRPS子单元的可用性.
- 研究生物过程不同阶段NRPS可用性的动态变化.
主要方法:
- 通过将NRPS子单元 (SrfAA-SrfAD) 与用于光标记的fefp基因融合,构建了四种Bacillus subtilis传感器菌株 (BMV25-BMV28).
- 使用FPCountR包监测光信号 (RFU) 和转换为相对NRPS分子数.
- 传感器菌株与父菌株 (BMV9) 之间的生长和表面蛋白生产率的比较.
主要成果:
- 传感器菌株的生长和表面活性蛋白的生产率与父菌株相似.
- 光信号显示了各种菌株的NRPS可用性,BMV27显示了最高的RFU.
- 在指数增长结束时,NRPS可用性达到顶峰,并在静止阶段下降.
结论:
- 开发的基于光的系统能够有效地在体内定量NRPS动态.
- NRPS的可用性受到生长阶段的影响,这表明在营养有限的条件下进行调节.
- 这种定量框架支持优化表面素生产和其他非核糖体生物工艺.
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