核糖体池工程增加了蛋白质生物合成产量
Camila Kofman1, Jessica A Willi1, Ashty S Karim1
1Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208, United States.
ACS central science
|April 29, 2024
概括
细菌核糖体具有影响蛋白质生产的序列变化. 通过使用特定的核糖体RNA (rRNA) 操作子来设计这些核糖体,可以显著提高合成生物学和生物制造的蛋白质产量.
科学领域:
- 分子生物学分子生物学
- 合成生物学 合成生物学
- 细菌遗传学 细菌遗传学
背景情况:
- 细菌核糖体对于蛋白质合成至关重要,具有多样化的核糖体RNA (rRNA) 操作序列.
- 由于这些本源序列变异而产生的功能差异尚未得到充分理解.
- 了解这些差异是优化合成生物学应用中的核糖体的关键.
研究的目的:
- 为了研究 *Escherichia coli* rRNA操作子中的序列多态性对核糖体性能的功能影响.
- 评估由不同rRNA操作子组合衍生的核糖体的蛋白质生产能力.
- 探索工程核糖体池的潜力,以提高蛋白质生物制造产量.
主要方法:
- 利用*体外*的核糖体合成和翻译平台,分析来自七个*大肠杆菌*的rRNA操作子中的16S和23SrRNA独特组合的核糖体.
- 产生表达单个rRNA操作子的细菌菌株,用于*in vivo*分析.
- 使用医学和工业相关蛋白质组的评估蛋白质生产产量.
主要成果:
- 原生 *E. coli* rRNA 操作子的序列变化显著改变了核糖体功能和蛋白质生产效率.
- 与野生类型异质池相比,一些纯化*in vivo*表达的同质核糖体池表现出优异的性能.
- 一个仅表达操作子A的细胞溶解体的核糖体显示了对标蛋白的实质性产量增加.
结论:
- 原生rRNA操作子序列多态性直接影响核糖体功能和蛋白质合成.
- 通过选择特定的rRNA操作子来设计核糖体池,可以提高生物制造中的蛋白质产量.
- 核糖体池工程为优化蛋白质生产和理解核糖体进化提供了一种新的策略.
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