A serine-substituted P450 catalyzes highly efficient carbene transfer to olefins in vivo.

A serine-substituted P450 catalyzes highly efficient carbene transfer to olefins in vivo.
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DOI:
10.1038/nchembio.1278
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发表时间:
2013-08
影响因子:
14.8
通讯作者:
Brustad EM
Brustad EM
中科院分区:
生物学1区
文献类型:
--
作者:
Coelho PS;Wang ZJ;Ener ME;Baril SA;Kannan A;Arnold FH;Brustad EM

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非天然化学反应的基因编码催化剂将为化学品的可持续生产开辟新的途径。我们设计了一个独特的丝氨酸血红素连接细胞色素“P411”,催化有效和选择性的卡宾转移从重氮酯烯烃在完整的大肠杆菌细胞。细胞色素P450 BM 3中的突变C400 S在411 nm处产生特征性亚铁-CO Soret峰,消除单加氧活性,提高静息状态Fe III/II还原电位,并显著提高NAD(P)H驱动的环丙烷化活性。
Genetically encoded catalysts for non-natural chemical reactions will open new routes to sustainable production of chemicals. We designed a unique serine-heme ligated cytochrome “P411” that catalyzes efficient and selective carbene transfers from diazoesters to olefins in intact Escherichia coli cells. The mutation C400S in cytochrome P450BM3 gives a signature ferrous-CO Soret peak at 411 nm, abolishes monooxygenation activity, raises the resting state FeIII/II reduction potential, and significantly improves NAD(P)H-driven cyclopropanation activity.
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