Engineering Cytochrome P450s for Enantioselective Cyclopropenation of Internal Alkynes
Engineering Cytochrome P450s for Enantioselective Cyclopropenation of Internal Alkynes
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DOI：
10.1021/jacs.0c01313
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发表时间：
2020-04-15
影响因子：
15
通讯作者：
Arnold, Frances H.
中科院分区：
文献类型：
--
作者：
Chen, Kai;Arnold, Frances H.
我们报告了一个工程化细胞色素P450酶的生物催化平台，通过卡宾转移到内部炔来进行有效的环丙烯合成。丝氨酸连接的P450变体P411-C10的定向进化产生了一系列工程化的P411酶，这些酶一起容纳多种内部芳香族炔作为环丙烯化的底物，具有前所未有的效率和立体选择性（高达5760 TTN，并且全部具有> 99.9%ee）。使用带有炔丙基醚基团的内部脂肪族炔，不同的P411变体可以选择性地催化环丙烯形成、卡宾插入炔丙基C-H键或[3 + 2]-环加成。这种可调的反应选择性进一步突出了使用基因编码催化剂来解决化学选择性挑战的好处。
We report a biocatalytic platform of engineered cytochrome P450 enzymes to carry out efficient cyclopropene synthesis via carbene transfer to internal alkynes. Directed evolution of a serine-ligated P450 variant, P411-C10, yielded a lineage of engineered P411 enzymes that together accommodate a variety of internal aromatic alkynes as substrates for cyclopropenation with unprecedented efficiencies and stereoselectivities (up to 5760 TTN, and all with >99.9% ee). Using an internal aliphatic alkyne bearing a propargylic ether group, different P411 variants can selectively catalyze cyclopropene formation, carbene insertion into a propargylic C-H bond or [3 + 2]-cycloaddition. This tunable reaction selectivity further highlights the benefit of using genetically encoded catalysts to address chemoselectivity challenges.