在扩展模拟过程中,工程Escherichia coli Nissle 1917菌株的遗传异质性
Lara P Munkler1, Elsayed T Mohamed1, Ruben Vazquez-Uribe2
1Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Lyngby, Denmark.
Metabolic engineering
|August 7, 2024
概括
工程微生物的遗传不稳定性是微生物组治疗的挑战. 删除易发生错误的DNA聚合酶减少了突变,但没有防止基因中断,需要进一步的稳定性策略.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 基因工程是一种基因工程.
背景情况:
- 先进的微生物组治疗利用工程微生物来治疗疾病.
- 工程微生物的遗传不稳定性是已知的生物制造挑战,但在微生物组疗法中尚未研究.
研究的目的:
- 为了研究工程Escherichia coli Nissle 1917菌株的遗传不稳定性,生产治疗性蛋白质.
- 评估删除易出错的DNA聚合酶 (polB,dinB,umuDC) 对应变稳定性的影响.
主要方法:
- 工程化大肠杆菌尼斯尔1917菌株产生干白素2和阿尔达费明的菌株在150代的时间里进化了.
- 使用自动化平台进行了高通量微生物进化.
- 整个基因组短读测序被用来分析遗传突变.
主要成果:
- 在六个不同的菌株和五个独立的血统中观察到基于pmut的生产等离子体的显著遗传不稳定性.
- 删除容易出错的DNA聚合酶可以使突变率降低12%-43%.
- 尽管突变减少了,但生产相关基因的中断仍然存在.
结论:
- 遗传不稳定性仍然是先进微生物组治疗的关键问题.
- 虽然减少易出错的聚合酶提供了部分稳定性,但不足以防止关键基因中断.
- 需要额外的策略来确保这些治疗方法的长期稳定性和有效性.
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