Major
Forensic Science
Access Type
Open Access
Abstract
Β-glucosidase is an enzyme that is essential for the hydrolysis of cellulase and is responsible for glucose production. Many mutations can occur within this enzyme, each has different enzymatic effects and likelihood of expression. For this experiment, enzymatic data was collected regarding the mutation BglB (Y118F) and compared to the wild-type data--which does not contain a mutation. This was done by designing the protein through the Foldit program, sequencing it, transforming it, purifying, expressing, and analyzing it. The analysis of this mutation consisted of a kinetic enzymatic assay and SDS PAGE. The BglB Y118F mutation was expressed, but not purified because of the absence of a band in the BglB Y118F lane on the SDS-PAGE gel. After performing the enzymatic assays, the resulting data was submitted to the D2D database, and various data (including Km, Vmax, and kcat/Km) were generated. The resulting data further proved that the BglB Y118F mutation showed better catalytic efficiency and substrate binding than the wild type.
Community Partners
Ashley Vater and the Justin Siegal Lab at UC-Davis
Faculty Mentors & Instructors
Emma Feeney, PhD, Biology Department
Supported By
The National Science Foundation
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Protein Purification and Expression of β-Glucosidase Mutant Y118F
Β-glucosidase is an enzyme that is essential for the hydrolysis of cellulase and is responsible for glucose production. Many mutations can occur within this enzyme, each has different enzymatic effects and likelihood of expression. For this experiment, enzymatic data was collected regarding the mutation BglB (Y118F) and compared to the wild-type data--which does not contain a mutation. This was done by designing the protein through the Foldit program, sequencing it, transforming it, purifying, expressing, and analyzing it. The analysis of this mutation consisted of a kinetic enzymatic assay and SDS PAGE. The BglB Y118F mutation was expressed, but not purified because of the absence of a band in the BglB Y118F lane on the SDS-PAGE gel. After performing the enzymatic assays, the resulting data was submitted to the D2D database, and various data (including Km, Vmax, and kcat/Km) were generated. The resulting data further proved that the BglB Y118F mutation showed better catalytic efficiency and substrate binding than the wild type.