苏氨酸醛缩酶的结构与功能及其在药物合成中的应用  

Structure and Function of Threonine Aldolase and Its Application in Pharmaceutical Synthesis

作　　者：何远志 冯雁[1] HE Yuan-Zhi;FENG Yan(School of Life Sciences and Biotechnology,State Key Laboratory of Microbial Metabolism,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学生命科学技术学院,微生物代谢国家重点实验室,上海200240

出　　处：《生物化学与生物物理进展》2023年第5期962-977,共16页Progress In Biochemistry and Biophysics

基　　金：国家重点研发计划(2020YFA0907700);国家自然科学基金(32271306)资助项目。

摘　　要：β-羟基-α-氨基酸(β-hydroxy-α-amnio acids,HAAs)是一类广泛应用于制药工业的重要手性中间体。由于其含有双手性中心(C_(α)和C_(β)),探索其严格立体选择性的生物合成方法备受关注。苏氨酸醛缩酶(threonine aldolase,TA)可在温和条件下催化不同类型的醛与氨基酸缩合构筑丰富的HAAs产物库,显示了工业应用潜力。由于目前表征的TA普遍存在对Cβ立体选择性不严格、活性较低以及催化机制不清晰等问题,为其在HAAs合成中的应用带来了挑战。本文综述了TA在新酶挖掘、结构与催化机理解析、蛋白质工程以及合成应用等方面的研究进展,为推动酶催化绿色、高效合成手性药物提供参考。β-Hydroxy-α-amino acids(HAAs)are a class of important chiral intermediates and have a wide range of uses in the pharmaceutical industry.Due to its adjacent chiral centers,it has attracted much attention to exploring strictly stereoselective biosynthesis methods.Threonine aldolase(TA)can catalyze the aldol reaction in one step for the synthesis of HAAs under mild conditions.TAs also exhibit a broad substrate spectrum and can catalyze the condensation of various aldehydes and amino acids,thereby constructing rich HAA libraries with immense potential for industrial applications.In this review,we have summarized the research progress of TA,including new enzyme mining,structure,catalytic mechanism analysis,high-throughput screening methods,protein engineering,and synthesis applications.Notably,we mainly focus on the research achievements of TA in structure-function relationship,mechanism analysis,and protein engineering.Currently,the catalytic process of TA has been elucidated,where it catalyzes the aldol reaction through the Schiff base exchange mechanism.Additionally,researchers have proposed diastereoselectivity regulating mechanisms of TA such as the“pathway hypothesis”and“dual conformation hypothesis”,which provide a foundation for unraveling the mystery of TA diastereoselectivity regulation.Moreover,significant progress has been made in TA molecular evolution,with multiple mutants obtained that showed strict diastereoselectivity synthesis for various chiral HAAs and their intermediates.Furthermore,we have discussed the current challenges and prospects of TA,which will guide for accelerating the industrial application of TAs as tool enzymes for synthesizing high-value HAAs compounds.

关 键 词：苏氨酸醛缩酶 β-羟基-α-氨基酸 晶体结构 立体选择性 蛋白质工程 

分 类 号：Q5[生物学—生物化学] Q7