Major
Biology
Anticipated Graduation Year
2027
Access Type
Restricted Access
Abstract
This study examined how a single amino acid mutation (W185R) affects the structure and function of the β-glucosidase (BglB) enzyme from Paenibacillus polymyxa. Using protein modeling, we predicted that the mutation would disrupt folding and reduce enzyme stability. The mutated gene was confirmed through plasmid preparation and sequencing, then transformed into E. coli for protein expression. The protein was purified and analyzed using SDS-PAGE and Western blot. Enzymatic activity was tested using a kinetic assay. Results suggest that the mutation decreased enzyme function, likely due to structural changes, supporting our hypothesis that W185R negatively impacts catalytic efficiency.
Faculty Mentors & Instructors
Dr Emma Feeney, Senior Lecturer, Biology Department
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Site-Directed Mutagenesis of BlgB (W185R) and Its Impact on Recombinant Expression and Enzymatic Function
This study examined how a single amino acid mutation (W185R) affects the structure and function of the β-glucosidase (BglB) enzyme from Paenibacillus polymyxa. Using protein modeling, we predicted that the mutation would disrupt folding and reduce enzyme stability. The mutated gene was confirmed through plasmid preparation and sequencing, then transformed into E. coli for protein expression. The protein was purified and analyzed using SDS-PAGE and Western blot. Enzymatic activity was tested using a kinetic assay. Results suggest that the mutation decreased enzyme function, likely due to structural changes, supporting our hypothesis that W185R negatively impacts catalytic efficiency.