Major

Forensic Science

Anticipated Graduation Year

2024

Access Type

Open Access

Abstract

The BglB enzyme, B-glucosidase, is a component in cellulase. It plays a role in catalyzing the enzymatic reaction of glucose formation during cellulose hydrolysis (Wright RM et al, 1992). We performed a mutation (M114S) on the BglB enzyme and performed data analysis to characterize the mutation's effects on the protein’s structure and function. The series of laboratory methods performed included sequencing, bacterial transformation, affinity purification, enzymatic kinetic assay, and finally analyzing our data with unsuccessful submission to the D2D database. We hypothesized that our BglB mutation, M114S, would show decreased catalytic efficiency compared to the wild type. We confirmed that our mutant had lower enzymatic activity based on the assay data and Michaelis-Menten plot showing little to no activity above background noise for increasing substrate concentrations.

Community Partners

Justin Siegel and Ashley Vater from UC Davis

Faculty Mentors & Instructors

Dr. Emma Feeney

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

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Analyzing the Impact of the BglB (M114S) Mutation on Catalytic Efficiency

The BglB enzyme, B-glucosidase, is a component in cellulase. It plays a role in catalyzing the enzymatic reaction of glucose formation during cellulose hydrolysis (Wright RM et al, 1992). We performed a mutation (M114S) on the BglB enzyme and performed data analysis to characterize the mutation's effects on the protein’s structure and function. The series of laboratory methods performed included sequencing, bacterial transformation, affinity purification, enzymatic kinetic assay, and finally analyzing our data with unsuccessful submission to the D2D database. We hypothesized that our BglB mutation, M114S, would show decreased catalytic efficiency compared to the wild type. We confirmed that our mutant had lower enzymatic activity based on the assay data and Michaelis-Menten plot showing little to no activity above background noise for increasing substrate concentrations.