Major
Neuroscience
Anticipated Graduation Year
2026
Access Type
Open Access
Abstract
This study characterized the β-glucosidase B (BglB) mutant P41L within the Design2Data (D2D) framework. We hypothesized that substituting proline with leucine at position 41, a non-active-site residue, would maintain catalytic efficiency (kcat/Km) but decrease thermal stability (T50) relative to wild-type (WT) BglB. Foldit modeling, plasmid sequencing, expression in E. Coli, and affinity purification were used to validate the construct. Kinetic parameters with p-nitrophenyl-β-D-glucopyranoside and T50 values across 30–50 °C were obtained via plate-reader assays and D2D fits. P41L displayed higher catalytic efficiency and a T50 comparable to WT, refuting our hypothesis and suggesting an efficiency-enhancing, stability-neutral mutation.
Community Partners
Ashley Vater and Justin Siegel Lab at UC Davis, Dr. Emma Feeney PhD at Loyola University Chicago
Faculty Mentors & Instructors
Dr. Emma Feeney PhD at Loyola University Chicago
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
β-glucosidase B Mutant P41L Characterization and Analysis
This study characterized the β-glucosidase B (BglB) mutant P41L within the Design2Data (D2D) framework. We hypothesized that substituting proline with leucine at position 41, a non-active-site residue, would maintain catalytic efficiency (kcat/Km) but decrease thermal stability (T50) relative to wild-type (WT) BglB. Foldit modeling, plasmid sequencing, expression in E. Coli, and affinity purification were used to validate the construct. Kinetic parameters with p-nitrophenyl-β-D-glucopyranoside and T50 values across 30–50 °C were obtained via plate-reader assays and D2D fits. P41L displayed higher catalytic efficiency and a T50 comparable to WT, refuting our hypothesis and suggesting an efficiency-enhancing, stability-neutral mutation.