New enzyme-catalyzed reactions from catalytically promiscuous ancestral enzymes

来自明尼苏达大学双子城分校的Romas Kazlauskas博士和Antony Dean博士获得了化学、生物工程、环境和运输系统部门的生命过程化学计划和化学、生物工程、环境和运输系统部门的生物技术、生化和生物质工程计划颁发的这一奖项，他们将利用基因合成重振祖先的通用酶。目标是羟腈裂解酶的祖先，它催化碳-碳键的形成：氰化物与醛的加成。这项研究将测试这些祖先酶催化其他碳-碳键形成反应的能力，并针对不同的反应进行优化。目前化学合成中的一个挑战是利用环境友好的条件和试剂来催化和立体选择性地组装复杂的分子。生物催化是一种潜在的解决方案，但目前生物催化剂的反应范围有限。这项工作构成了一种新的方法来发现催化非自然反应的生物催化剂，它使用碳-碳键形成反应作为测试案例。对复活的祖先酶的研究试图回答新的酶和代谢途径如何进化的基本问题。此外，复活的祖先酶可能是设计新酶的起点，因为祖先酶可以催化额外的化学反应，包括潜在有用的非自然反应。拟议的工作将培训本科生、研究生和博士后，以化学方法解决跨学科问题，积极招收和接纳化学科学中代表性不足的群体的学生，并支持向小学生进行外联。

In this award from the Chemistry of Life Processes Program in the Division of Chemistry and the Biotechnology, Biochemical, and Biomass Engineering Program in the Chemical, Bioengineering, Environmental, and Transport Systems Division, Drs. Romas Kazlauskas and Antony Dean, from the University of Minnesota, Twin Cities, will resurrect ancestral generalist enzymes using gene synthesis. The targets are ancestors of hydroxynitrile lyases, which catalyze a carbon-carbon bond formation: addition of cyanide to aldehydes. The research will test the ability of these ancestral enzymes to catalyze other carbon-carbon bond forming reactions and optimize them for different reactions. A current challenge in chemical synthesis is to assemble complex molecules catalytically, and stereoselectively while using environmentally friendly conditions and reagents. Biocatalysis is a potential solution, but the reaction range of current biocatalysts is limited. This work constitutes a new approach to discover biocatalysts that catalyze unnatural reactions, which uses carbon-carbon bond forming reactions as a test case. Studies of resurrected ancestral enzymes seek to answer basic questions of how new enzymes and metabolic pathways evolve. In addition, resurrected ancestral enzymes may be starting points for engineering new enzymes because ancestral enzymes can catalyze additional chemical reactions, including potentially useful unnatural reactions. The proposed work will train under-graduate, graduate and postdoctoral students in a chemical approach to solving interdisciplinary problems, actively recruit and include students from groups underrepresented in chemical sci-ences, and support outreach to elementary school students.