Major
Forensic Science
Anticipated Graduation Year
2024
Access Type
Open Access
Abstract
This presentation will describe the experience of generating a mutant enzyme, characterizing its function, and making protein function predictions. This project began by designing the H328N mutation for our selected protein, β–glucosidase B (BglB), using the molecular modeling tool, Foldit. Following preparation of plasmid DNA containing H328N, DNA sequencing was performed prior to expression and purification. An enzyme kinetic assay was conducted to facilitate characterization of activity. Lastly, the purity was analyzed using SDS-PAGE. We utilized our calculated system energy values obtained from the Foldit models in conjunction with examination of intermolecular interactions to predict the outcome of our experiment.
Faculty Mentors & Instructors
Emma Feeney, PhD, Department of Biology
Supported By
Justin Siegel, PhD; Ashley Vater, MS; National Science Foundation
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Determining the Catalytic Efficiency of β-Glucosidase B (BglB) H328N Mutant
This presentation will describe the experience of generating a mutant enzyme, characterizing its function, and making protein function predictions. This project began by designing the H328N mutation for our selected protein, β–glucosidase B (BglB), using the molecular modeling tool, Foldit. Following preparation of plasmid DNA containing H328N, DNA sequencing was performed prior to expression and purification. An enzyme kinetic assay was conducted to facilitate characterization of activity. Lastly, the purity was analyzed using SDS-PAGE. We utilized our calculated system energy values obtained from the Foldit models in conjunction with examination of intermolecular interactions to predict the outcome of our experiment.