32nd Annual Student Research Conference
Disulfide Bond Forming Enzyme Expression and Purification
Kerstin E. Peterson
Dr. Cassidy Dobson, Faculty Mentor
Disulfide Bond Forming Enzyme (DBF), a unique and interesting chaperone protein found in the hyperthermophilic bacteria Sulfolobus solfataricus. DBF has the ability to chaperone disulfide bonds without the presence of cysteine unlike all other disulfide chaperones implying a novel mechanism of action. In order to elucidate the mechanism of DBF, pure, concentrated protein is required. DBF is overexpressed in E. coli using the induction sugar rhamnose, purified by immobilized affinity column chromatography and analyzed via SDS-PAGE. To increase purity and concentration, ammonium sulfate precipitation was performed and evaluated by SDS-PAGE. The ability to purify DBF through ammonium sulfate leads to an increase in purification efficiency, and further testing can then be used to determine the mechanism of chaperone activity of DBF.
Keywords: disulfide, cysteine, expression, purification, ammonium sulfate, E. coli
Topic(s):Biochemistry and Molecular Biology
Chemistry
Presentation Type: Oral Paper
Session: 111-5
Location: MC 212
Time: 9:30