Design to Data for Mutant of β-Glucosidase B from Paenibacillus polymyxa : G23S
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β-glucosidase (BglB) from Paenibacillus polymyxa was mutated (G23S, Rosetta/Foldit numbering; G26S, conventional numbering) to assess structural and functional changes. Foldit modeling and prior Design 2 Data (D2D) database results led us to hypothesize that this mutation would increase substrate binding affinity and catalytic efficiency, with a moderate reduction in thermal stability. The mutant protein was expressed, purified, and analyzed using kinetics and thermal stability assays. Relative to the wild-type (WT), G23S exhibited a similar binding affinity (similar K m ), an approximately 2-fold increase in turnover number (k cat ) and catalytic efficiency (k cat /K m ), an almost 14-fold increase in maximum reaction velocity (V max ) and a slight decrease in thermostability (T 50 ). The results largely support the hypothesis, indicating that changes in residue 23 can enhance catalytic power while minimally compromising stability.
