酶促立体可调控一步合成双手性中心化合物方法及其调控机理

发展选择性更好的酶促方法，特别是能够有效地调控选择性、并能一步合成具有多手性中心化合物的方法，对于复杂有机手性合成而言至关重要。项目主要通过对CALB脂肪酶的酸和醇结合口袋进行蛋白质工程改造，构建同时对酸和醇都具有高选择性的活性位，实现一步拆分醇和酸两个手性中心，制备单一构型的双手性中心化合物；并通过对酸和醇底物选择性的不同组合，有效地实现在4种可能的产物构型间自由调控；探索CALB脂肪酶结构与立体选择性之间的关系，深入认识酶结构对催化立体选择性的影响规律；建立酸和醇同时动态动力学拆分的方法，并利用该方法合成重要的双手性中心药物分子。本项目的研究有助于建立更加高效、绿色和原子经济性的酶促合成新方法，为制药工业和精细化工领域的立体选择性合成方法提供更好的酶催化剂积累基础，在生物催化和有机合成相关领域有重要的应用前景。

It is very important for stereoselective complex organic synthesis to develop enzymatic synthesis strategy with higher and easily controllable stereoselectivity, which can be used for one-step synthesis of optically pure compounds with several chiral centers. This project mainly focuses on the rational redesign of the alcohol binding pocket and acyl binding pocket of CALB, and attempts to construct CALB mutants with high stereoselectivity for both alcohol and acid substrates, and then will use these mutated CALB for the synthesis of optically pure compounds with double chiral centers. It is possible to control the stereoselectivity for the four possible enantiomers respectively, through the suitable combination between the two selectivity of CALB for alcohol and acid substrate. This project also plans to explore the relationship between the structure of CALB mutants and their stereoselectivity, and provide some knowledge about how the mutated amino acids influence the selectivity of CALB. Furthermore, simultaneous dynamic kinetic resolution of alcohol and acid moieties of the ester with two chiral centers will be investigated, and used for the synthesis of an important chiral drug model. The obtained results of this project will be useful for the development of more efficient, more atom economical and greener enzymatic synthesis, and have important application potentials in the fields of biocatalysis and organic synthesis.