Docking Studies and Molecular Cloning of Alpha/Beta Hydrolase from Novel Streptomyces werraensis KN23 for Enhanced Keratinase Activity

Abstract

Alpha/beta hydrolases constitute an expansive functional group of enzymes present in bacteria, archaea, and eukaryotes. The present study demonstrates the gene encoding alpha/beta hydrolase (consisting of 1596 bp nucleotides) of novel Streptomyces werraensis KN23, the sample was subjected to isolation, sequencing, and then uploaded to the NCBI GenBank database with the accession number OQ511280. Validation of the modeled protein's 3D structure was conducted through Ramachandran's plot, revealing
that, for the template proteins S. werraensis FS97 and KN23, 92.51% and 97.48%, respectively, of the 532 the residues were placed in the area with the highest preference. Docking studies revealed optimal binding affinities of the alpha/beta hydrolase with beta keratin, of values -259.20 kcal/mol and -227.70 kcal/mol for the template strains S. werraensis FS97 and S. werraensis KN23, respectively. Expression of the S. werraensis KN23 alpha/beta hydrolase gene, the sample had a keratinase activity of 51.60 U/ml. The binding affinities between the deduced protein and beta keratin were calculated using computational methods. The alpha/beta hydrolase gene was transcribed and translated in E. coli DH5α, leading to a substantial increase in keratinase activity (93.38 U/ml).