Major
Forensic Science
Anticipated Graduation Year
2025
Access Type
Open Access
Abstract
This experiment's purpose was to characterize the Bgl B mutant (Y118F) by its enzyme kinetics. The wild-type Bgl B and the Y118F mutant were compared in Foldit, a chemical structure modeling software, and it was determined that our mutation would see a decrease in catalytic efficiency, substrate binding ability, and overall effectiveness. The experiment was completed with the goal of gathering data to submit to Design2Data a protein modeling algorithm. The experimentation entailed preparing the plasmid, expressing and purifying the mutant protein, analyzing the mutant using a kinetics assay and SDS-PAGE, and then interpreting and visualizing the results. The results of the SDS-PAGE did not provide much use due to little visible banding present, indicating that there are low concentrations of the mutant present. The kinetic assay results support the Kcat decreasing for the mutant but results were vague and inconclusive for the Km.
Community Partners
Ashley Vader and the Justin Seigel Laboratory at UC-Davis, National Science Foundation
Faculty Mentors & Instructors
Emma Feeney Ph.D.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Kinetic Efficiency Determination of Mutant Enzyme Bgl B (Y118F)
This experiment's purpose was to characterize the Bgl B mutant (Y118F) by its enzyme kinetics. The wild-type Bgl B and the Y118F mutant were compared in Foldit, a chemical structure modeling software, and it was determined that our mutation would see a decrease in catalytic efficiency, substrate binding ability, and overall effectiveness. The experiment was completed with the goal of gathering data to submit to Design2Data a protein modeling algorithm. The experimentation entailed preparing the plasmid, expressing and purifying the mutant protein, analyzing the mutant using a kinetics assay and SDS-PAGE, and then interpreting and visualizing the results. The results of the SDS-PAGE did not provide much use due to little visible banding present, indicating that there are low concentrations of the mutant present. The kinetic assay results support the Kcat decreasing for the mutant but results were vague and inconclusive for the Km.