在蛋白质中功能性替换histidine以产生非正规的氨基酸依赖生物
Fei Gan1, Renhe Liu1, Feng Wang1
1California Institute for Biomedical Research (Calibr) , La Jolla , California 92037 , United States.
Journal of the American Chemical Society
|March 7, 2018
概括
研究人员开发了一种方法来创造需要特定非正规氨基酸生长的微生物,从而提高了疫苗和生物制的安全性. 这涉及到用功能模拟物取代必需的histidine氨基酸.
科学领域:
- 合成生物学 合成生物学
- 生物技术是生物技术.
- 微生物遗传学微生物遗传学
背景情况:
- 需要非正规氨基酸的生物对活疫苗和生物制至关重要.
- 对必不可少的氨基酸残留物的基因操纵提供了一种途径来实现辅助性.
研究的目的:
- 开发一种简单的策略来创造具有严格成长依赖于非正规氨基酸的有机体.
- 为了证明金属酶中一个关键的胺残留物的功能替代,用一个非正规的类似物.
主要方法:
- 在含的酶曼诺-6-异相酶 (ManA) 中,基因替换了histidine (His) 残留物,用3-methyl-l-histidine (MeH) 来代替.
- 设计一种大肠杆菌菌株与一个进化的ManA变体 (Opt5) 结合N262S和H264MeH替代品.
- 评估工程化大肠杆菌菌株的生长依赖性和逆转率.
主要成果:
- 在ManA.中,3-甲基-l-histidine (MeH) 功能地替代了一个关键的金属结合联体 (H264) .
- 工程ManA变体 (Opt5) 显示了与野生类型酶相当的活性.
- 工程化大肠杆菌菌株表现出严格依赖MeH的生长,反转率非常低.
结论:
- 这种策略提供了一种强大的方法来产生辅食性生物.
- 这种方法可能适用于其他含有必要的胺残留物的金属或非金属蛋白.
- 这种方法提高了重组生物和活疫苗的生物制和安全性.
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