Catalyst Discovery with Engineered Thiamin-Dependent Transketolase: Cofactor Catalysis

纽约州立大学布法罗分校化学学部的启动基金资助了Steven T. Diver教授的研究。本研究的目的是通过体内筛选方法鉴定新的催化剂。该酶体系为转酮醇酶，其催化机制为硫胺焦磷酸辅因子。通过酵母转酮醇酶基因的突变，开发了一个系统，在该系统中，酵母只有在通过结合新的辅因子恢复催化功能时才能生长。设计、合成了各种硫胺素类似物，并测试了它们作为新的催化辅助因子的活性。生长表型用于发现新的辅因子催化剂作为体内选择策略。本研究探索了一种新型催化剂发现的新方法，利用活细胞来传达反应的价值。这项工作加强了对酶系统结构和功能之间关系的理解。

The starter grant award of the Chemistry Division to the State University of New York at Buffalo supports the research of Professor Steven T. Diver. The objective of the research is to identify new catalysts by an in vivo screening method. The enzyme system is transketolase with thiamin pyrophosphate cofactor as its catalytic machinery. Through mutation of the transketolase gene in yeast, a system is developed in which yeast grow only when catalytic function is recovered by incorporation of a new cofactor. Various thiamin analogs are designed, synthesized, and tested for activity as new catalytic cofactors. The growth phenotype is used to discover a new cofactor catalyst as an in vivo selection strategy. The research explores a new method for novel catalyst discovery, using a living cell to communicate a reaction's worthiness. The work enhances understanding of the relationship between structure and function in enzyme systems.