Centennial Honors College Western Illinois University Undergraduate Research Day 2014 Poster Presentation Cloning of a Putative L-3-hydroxyacyl-CoA Dehydrogenase from Micrococcus Luteus WIUJH20 Antoinee Watson-Taylor Faculty Mentors: Lisa Wen and Jenq-Kuen Huang Chemistry Enzymes are biological catalysts that carry out chemical reactions in all living organisms. Thus, enzymes play important roles in maintaining health and vitality. The present research is aimed at understanding how enzyme works by studying two enzymes: secondary alcohol dehydrogenase (2°-ADH) and L-3-hydroxyl-CoA dehydrogenase (L-3-HAD). Alcohol dehydrogenases are enzymes that break down alcohols and catalyze interconversion between alcohols and aldehydes or ketones. L-3HADs are enzymes involved in the fatty acid oxidation pathway. Through the amino acid sequences alignment study of 2°-ADH from Micrococcus luteus WIUJH20 (M. luteusWIUJH20) with those in the Protein Database, it was discovered that the 2°-ADH has 89% sequence identity with the annotated L-3-HAD from M. luteus NCTC2665. However, we have ruled out that the annotated L-3-HAD doesn’t belong to L-3-HADs family by kinetics studies of the purified 2°-ADH. The two enzymes are different in their selection of reactants/substrates. Therefore, the L-3-HAD and 2°-ADH are excellent model systems for studying structure-function relationship of the enzymes and probing the amino acid residues within the enzymes that are responsible for achieving such specificity. Such study requires production of large amounts of each enzyme obtained from M. luteusWIUJH20 to make comparison significant. The short-term objective of this research is to isolate the gene encoding L-3-HAD from M. luteus WIUJH20 by molecular cloning approach to produce large quantity of L-3HAD. The long-term goal of this project is to understand how enzymes react specifically to their substrates. This poster presentation will include strategies for the gene cloning and preliminary results.