Catalyst Discovery with Engineered Thiamin-Dependent Transketolase: Cofactor Catalysis

化学系授予纽约州立大学布法罗分校的STARTER助学金支持史蒂文·T·迪弗教授的研究。本研究的目的是通过体内筛选的方法来寻找新的催化剂。该酶系为转酮醇酶，以硫胺素焦磷酸辅酶为催化机制。通过突变酵母中的转酮醇酶基因，建立了一种酵母只有在通过加入新的辅因子恢复催化功能的情况下才能生长的系统。各种硫胺素类似物被设计、合成，并作为新的催化辅因子进行活性测试。生长表型被用来发现一种新的辅因子催化剂作为体内选择策略。这项研究探索了一种发现新型催化剂的新方法，使用活细胞来传达反应的价值。这项工作加深了人们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.