Engineering the regioselectivity of Baeyer-Villiger monooxygenases

Emory University, Atlanta GA

Investigators

Abstract

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Stefan Lutz from Emory University to create novel biocatalysts for Baeyer-Villiger oxidation with enhanced regio-selectivity. Catalysts not only facilitate chemical reactions in industrial processes as diverse as producing petrochemical fuels and manufacturing pharmaceuticals, but they also govern the chemistry of life, playing a key role in driving biochemical reactions in cells. A better understanding of the fundamentals of (bio)catalysts, as well as advances in synthetic biology and biotechnology now allow scientists to manipulate and tailor biocatalysts for properties such as stability and recognition of new substances. These capabilities have created unprecedented opportunities to adapt these highly efficient and renewable catalysts for novel, unnatural functions needed in industrial applications, thereby creating new, safer and more sustainable processes. This project will provide graduate students with specialized training in preparing biocatalysts, as well as tailoring them to specific industrial applications. It also introduces undergraduate researchers, as well as high school teachers and students to the concepts of biocatalysis and green chemistry. The area of flavo-enzyme engineering for utility in synthetic chemistry is one that is ripe for development. Directed enzyme 'evolution' is a burgeoning field, but this enzyme class is relatively underexplored, and yet holds great potential for application to chemistry of interest. Employing a combination of flavin cofactor analog substitution and protein mutagenesis, native monooxygenases will be tailored for effective and selective conversion of asymmetric ketones to the corresponding ester regio-isomers. Lead candidates among these engineered enzymes will be characterized via comprehensive biochemical and biophysical analysis. These studies will provide detailed information concerning the impact of various engineering strategies and assist in developing accurate predictive frameworks for future enzyme redesigning endeavors.