19th Annual Student Research Conference
Science
Expression, Purification and Initial Characterization of Halobacterium Proline Dehydrogenase
Jeremy J. West
Dr. John J. Tanner (University of Missouri - Columbia), Dr. Russell G. Baughman, and Ms. Tommi A. White (University of Missouri - Columbia), Faculty Mentors
Nature recycles proline by converting it to glutamate. This 4-electron oxidation process is catalyzed by two catabolic enzymes, proline dehydrogenase (PRODH) and Δ1-pyrroline-5-carboxylate dehydrogenase (P5CDH). Inborn defects in PRODH and P5CDH result in the disorders hyperprolinemia I & II, respectively. The goal of this research is to characterize the structure and function of a newly discovered homologue of PRODH found in Archaea. Study of Archaea proteins may provide insights into homologous eukaryotic enzymes. The PRODH investigated here is from the Halobacterium (salt-loving). Preliminary results include testing the expression of Halobacterium PRODH (YusM) in two different E. coli expression systems, BL21(DE3)pLysS and Rosetta2. Parameters varied in these tests included time and temperature of induction as well as isopropyl-beta-D-thiogalactopyranoside concentration. Initial kinetic results also show improved activity after denaturing, and then renaturing the protein.
Keywords: protein, protein expression, protein purification, kinetics, proline, Halobacterium, denatured protein
Topic(s):Chemistry
Presentation Type: Oral Paper
Session: 41-3
Location: VH 1408
Time: 1:45