Engineering Carboxylic Acid Reductase for the Biosyntheses of Industrial Chemicals

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The structure of proteins determines their function. This project will attempt to modify several structural features of enzymes in order to obtain more productive functions and higher reaction rates. The enzymes are newly discovered and could help convert biomass into valuable chemical products very efficiently. The project will also provide interactive research experiences to students. The PIs will continue recruiting undergraduate and graduate students from underrepresented minorities to work on this project. The PIs will also be assembling and distributing an educational kit through the Nebraska EPSCoR Young Scientists Program, beginning with the HS level and expanding to the middle school level. This will provide these students with hands-on experience designing and creating different proteins.The proposed work focuses on the study and engineering of carboxylic acid reductases (CARs). The goal of the project is to expand the substrate specificity of CARs in order to efficiently reduce short-chain carboxylic acid metabolites into aldehydes. These serve as common and versatile biosynthetic intermediates that are readily to be further converted to alkanes, alkenes, alcohols, or amines. This proposal specifically targets CAR mutants that function on the reduction of three abundant carboxylic acid metabolites, including D- and L-lactic acids (C3 substrates), and glycolic acid (a two-carbon substrate). Further reduction of the aldehyde products leads to the formation of R- or S-1,2- propanediol and ethylene glycol, which are bulk industrial chemicals with broad applications, especially in the production of polyesters.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.