(493c) GDH to Adh: Complete Redesign of Glucose Dehydrogenase to Alcohol Dehydrogenase Conference: AIChE Annual MeetingYear: 2015Proceeding: 2015 AIChE Annual MeetingGroup: Food, Pharmaceutical & Bioengineering DivisionSession: Protein Structure, Function, and Stability Time: Wednesday, November 11, 2015 - 9:10am-9:30am Authors: Stahl, L., University of Stuttgart Bommarius, B., Georgia Institute of Technology Bommarius, A. S., Georgia Institute of Technology Pleiss, J., University of Stuttgart Tassoulas, L., Biocatalysts are increasingly used in industry to create enantiomerically pure compounds; routes employing dehydrogenases have been especially successful. Glucose dehydrogenase belongs to the extensive superfamily of SDRs (short chain dehydrogenase/reductase) and is currently a favored enzyme for cofactor regeneration. Glucose is its only substrate at a reasonable rate and the reaction is favored towards oxidation of glucose. A thermostable glucose dehydrogenase, previously developed at GeorgiaTech, was submitted to several rounds of site-directed mutagenesis, supported by database analysis and structure-guided design, to create an alcohol dehydrogenase based on a glucose dehydrogenase scaffold with a markedly changed substrate specificity. Key to the identification of the necessary amino acid hot spots within the sequence was a bioinformatics approach that involved an extensive data base analysis of existing SDRs using specific algorithms. The resulting 3-4 amino acid exchanges led to a complete change of substrate specificity away from glucose but respectable activity towards substrates such as cyclopentanol and 1,3-cyclohexandiol. By achieving the goal of de novo design of a glucose dehydrogenase, we demonstrate the successful implementation of bioinformatic tools and modeling that can accelerate development of novel substrate specificity in an otherwise limited enzyme system.