Characterization of Thermal Stability In Endoglucanases with Bioinformatics, Molecular Simulation, and Rosetta Design

The desire for increased protein thermal stability in commercial applications of cellulases motivates continued research focusing on the molecular aspects of protein stability.  Starting with a well-defined set of published data for mesophilic, thermophilic, and hyperthermophilic cellulase enzymes belonging to the Families 5 and 12 glycoside hydrolases, we have applied multiple techniques to gain insight into the structural features that confer thermal stability.  Bioinformatics, along with sequence and structural alignments, have been used as a preliminary means of identifying features relevant to thermal stability.  Rosetta simulations were then used to identify characteristic protein folding metrics in order to narrow down the critical structural features, which were subsequently evaluated by molecular dynamics.  Here, we present insights into the engineered molecular basis for increased thermal stability from molecular dynamics simulations of selected mutant structures compared to known thermal stable endoglucanases.