P450 Fingerprinting Method for Rapid Discovery of Terpene Hydroxylating P450 Catalysts with Diversified Regioselectivity

被引：99
作者：
Zhang, Kaidong [1 ]
El Damaty, Shady [1 ]
Fasan, Ruch [1 ]
机构：
[1] Univ Rochester, Dept Chem, Rochester, NY 14627 USA
来源：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2011年 / 133卷 / 10期
基金：
美国国家科学基金会;
关键词：
C-H BONDS; CYTOCHROME-P450; MONOOXYGENASE; ALKANE HYDROXYLATION; ELECTRON-TRANSFER; SUBSTRATE; OXIDATION; BINDING; CAMPHOR; SELECTIVITY; BACTERIAL;
D O I：
10.1021/ja109590h
中图分类号：
O6 [化学];
学科分类号：
0703 ;
摘要：
Engineered P450 enzymes constitute attractive catalysts for the selective oxidation of unactivated C-H bonds in complex molecules. A current bottleneck in the use of P450 catalysis for chemical synthesis is the time and effort required to identify the P450 variant(s) with the desired level of activity and selectivity. In this report, we describe a method to map the active site configuration of engineered P450 variants in high throughput using a set of semisynthetic chromogenic probes. Through analysis of the resulting 'fingerprints', reliable predictions can be made regarding the reactivity of these enzymes toward complex substrates structurally related to the fingerprint probes. In addition, fingerprint analysis offers a convenient and time-effective means to assess the regioselectivity properties of the fingerprinted P450s. The described approach can represent a valuable tool to expedite the discovery of P450 oxidation catalysts for the functionalization of relevant natural products such as members of the terpene family.
引用
收藏
页码：3242 / 3245
页数：4
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