College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, Jiangsu, 225009, China.
School of Life Science, Economic and Technology Development Zone, Anhui University, Hefei, Anhui, 230601, China.
Angew Chem Int Ed Engl. 2022 Jul 11;61(28):e202204290. doi: 10.1002/anie.202204290. Epub 2022 May 23.
Catalytic selective hydroxylation of unactivated aliphatic (sp ) C-H bonds without a directing group represents a formidable task for synthetic chemists. Through directed evolution of P450 hydroxylase, we realize oxyfunctionalization of unactivated C-H bonds in a broad spectrum of aliphatic carboxylic acids with varied chain lengths, functional groups and (hetero-)aromatic moieties in a highly chemo-, regio- and enantioselective fashion (>30 examples, Cβ/Cα>20 : 1, >99 % ee). The X-ray structure of the evolved variant, P450 -L78I/Q85H/G290I, in complex with palmitic acid well rationalizes the experimentally observed regio- and enantioselectivity, and also reveals a reduced catalytic pocket volume that accounts for the increased reactivity with smaller substrates. This work showcases the potential of employing a biocatalyst to enable a chemical transformation that is particularly challenging by chemical methods.
未活化脂肪族（sp ）C-H 键的催化选择性羟化对于合成化学家来说是一项艰巨的任务。通过 P450 羟化酶的定向进化，我们实现了在广泛的脂肪族羧酸中未活化 C-H 键的氧化官能化，这些羧酸具有不同的链长、官能团和（杂）芳基部分，具有高度的化学选择性、区域选择性和对映选择性（>30 个实例，Cβ/Cα>20 : 1，>99 %ee）。与棕榈酸形成复合物的进化变体 P450 -L78I/Q85H/G290I 的 X 射线结构很好地解释了实验观察到的区域和对映选择性，还揭示了催化口袋体积的减小，这解释了与较小底物反应性增加的原因。这项工作展示了利用生物催化剂实现化学转化的潜力，这种转化通过化学方法特别具有挑战性。
