34th Annual Student Research Conference
Expression and Purification of DBF
Corbin A. Estes
Dr. Cassidy Dobson, Faculty Mentor
Disulfide Bond Forming Enzyme (DBF) is a unique chaperone protein found in the hyperthermophile Sulfolobus solfataricus. DBF has the ability to chaperone disulfide bonds without the presence of cysteine, which separates it from other disulfide chaperones, implying a novel mechanism of action. DBF is overexpressed in E. coli, purified by immobilized metal affinity column chromatography, and analyzed via SDS-PAGE. One determines DBF activity by how well it can refold lysozyme. By measuring the activity of refolded lysozyme by UV-Vis spectroscopy, one can monitor the ability of DBF to correct disulfide bonds. In order to understand DBF’s unique mechanism, we must be able to study when and how DBF is able to refold disulfide interactions.
Keywords: DBF, Expression, Purification, Chaperone, Disulfide Bonds
Topic(s):Biochemistry and Molecular Biology
Chemistry
Biology
Presentation Type: Asynchronous Virtual Oral Presentation
Session: 3-6
Location: https://flipgrid.com/f86d186b
Time: 0:00