Cloning, sequencing and <em>in silico</em> analysis of β-glucosidase from <em>Bacillus subtilis</em> strain PS

Abstract

Glycosyl hydrolases are group of enzymes classified into 132 known families based on sequence similarity. Glycosyl hydrolase family 1 comprises of number of enzymes with a known activities including β-glucosidases, which have wide diversity and known to have important applications. In the present study, the β-glucosidase gene of glycosyl hydrolase family 1 from <em>Bacillus subtilis</em> strain PS was successfully cloned and expressed in <em>E. coli</em> BL21 (DE3) by using pET28 (a) vector system. Sequence characterization revealed a 1407 bp long nucleotide sequence with an open reading frame (ORF) encoding 469 amino acids and a calculated molecular mass of 51.6 kDa. Its translated protein sequence was used for comparative analysis to reference protein sequences of β-glucosidases from different organisms for multiple sequence alignment, phylogenetic tree construction, variation in biochemical features and distribution of motifs using various bioinformatics tools<em>.</em> The analysis revealed important sequence features and properties of this important family which would be helpful in further classification, diversity study and genetic engineering of glycosyl hydrolase family 1 β-glucosidases.