Exploring the Catalytic Significant Residues of Serine Protease Using Substrate-Enriched Residues and a Peptidase Inhibitor

Serine proteases are the most versatile proteolytic enzymes with tremendous applications in various industrial processes. This study was designed to investigate biochemical properties, critical residues, and catalytic potential of alkaline serine protease using in-silico approaches. The primary sequence analyzed ProtParam, SignalP, Phyre2 tools signal peptide, secondary structure, respectively. three-dimensional structure enzyme modeled MODELLER program present Discovery Studio followed by Molecular Dynamics simulation GROMACS 5.0.7 package CHARMM36m force field. measured performing docking casein- keratin-enriched while effect inhibitor studied phenylmethylsulfonyl fluoride, (PMSF) applying GOLDv5.2.2. weight, instability index, aliphatic isoelectric point for were 39.53 kDa, 27.79, 82.20 8.91, best model selected based on lowest MOLPDF score (1382.82) DOPE (-29984.07). analysis ProSA-web revealed a Z-score -9.7, whereas 88.86% residues occupied favored region Ramachandran plot. Ser327, Asp138, Asn261, Thr326 found as involved ligand binding execution biocatalysis. Our findings suggest that bioengineering these may enhance this enzyme.