用于选择性功能化循环化合物的P450碳酸转移酶的定向演化
Oliver F Brandenberg1, Kai Chen1, Frances H Arnold1
1Division of Chemistry and Chemical Engineering , California Institute of Technology , Pasadena , California 91125 , United States.
Journal of the American Chemical Society
|May 10, 2019
概括
研究人员开发了一种细胞P450酶,用于高效的碳转移反应,使异环和具有选择性功能. 这种生物催化剂为合成有价值的化学构件提供了金属催化剂的可持续替代品.
科学领域:
- 生物催化和酶工程
- 有机合成
- 医学化学
背景情况:
- 碳转移反应对于合成药和化学构件至关重要.
- 血蛋白碳酸转移酶提供可调节的活性位点以控制功能,但适合异环酶很少.
- 现有的酶的流量很低,或者需要昂贵的辅助因子.
研究的目的:
- 开发一种细胞P450酶,以有效地将碳转移到异环和循环.
- 开发一种高通量选方法,用于指导碳转移酶活性的演变.
- 扩大生物催化碳转移用于合成复杂分子的范围.
主要方法:
- 开发了一种光谱测量高通量选试验,用于1-甲基英多尔C3化.
- 使用定向进化,包括随机和有针对性的突变发生,以优化P450变体.
- 进行并行演化以增强醇化,酶选择性醇化和循环化活动.
主要成果:
- 进化了P450变体,具有11个氨基酸替代和删除的减少酶域,达到470次/分钟和18000次的总回转.
- 已经证明了醇的化学选择性C3化.
- 使用工程生物催化剂实现区域选择性醇化,反选择性醇化和循环烯循环化.
结论:
- 工程化血蛋白对于循环目标的选择性功能化是高度可调的.
- 定向进化是扩大新生酶催化剂能力的强大工具.
- 开发了高效的碳酸转移生物催化剂,为传统的金属催化剂提供可持续的替代品.
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