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Design to Data for mutants of β-glucosidase B from Paenibacillus polymyxa: Y333F, A88E, L219Q, A408H, Y173L, E340S, and Y422F

Maduros, A.; Farinsky, L.; Tagkopoulos, P.; Vater, A.; Siegel, J. B.

2026-02-05 biochemistry
10.64898/2026.02.04.703908 bioRxiv
Show abstract

This study explores computational design predictions related to experimental enzyme behavior by analyzing seven single-point mutants of {beta}-glucosidase B (BglB) from Paenibacillus polymyxa: Y333F, A88E, L219Q, A408H, Y173L, E340S, and Y422F. Each mutation was modeled using Foldit Standalone, and mutant selections were based on predicted thermodynamic stability changes of interest. Six of the seven mutants in this set yielded soluble, expressed protein. Most variants had similar catalytic efficiency compared to the wild type with one exception. The melting temperatures for most variants were also similar to the wild type. Correlation analysis revealed weak but potentially informative relationships between predicted {Delta}TSE and (a) thermal stability and (b) catalytic efficiency. These results further support known limitations of TSE score as a tool for single point mutation design and add to a growing dataset being generated to build the next generation of functionally predictive protein models.

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