Gene Fragments that encodes inulin hydrolysis enzyme from genomic Bacillus licheniformis: Isolation by PCR technique using new primers

Abstract

Fructose and Fructo-Oligosaccharides (FOS) are derived more practical from the enzymatic hydrolysis reaction of inulin. In this study, the gene fragments that encodes inulin hydrolysis enzyme were isolated from genomic B. licheniformis by Polymerase Chain Reaction (PCR) technique using new primers designed on conserved domain in family GH32 enzymes. Gene fragments were cloned into pGEM-T vector with E. coli as host cells and determined the sequence of nucleotide bases. Size of the gene fragment have been found 539 bp using the DPE.slF and DPE.eR primer pair. The gene fragment encodes 179 amino acid residues of protein fragment. The protein fragment has high homology with levanase of Bacillus subtilis BSn5, levanase of Bacillus subtilis subsp. subtilis str 168, exoinulinase of Pseudomonas mucidolents, exoinulinase of Paenibacillus polymyxa, exoinulinase of Geobacillus stearothermophilus, exoinulinase of Paenibacillus Aloe sp-11 and exoinulinase of Paenibacillus polymyxa SC2. The homology were 97, 97, 62, 62, 61, 62 and 62%, respectively. The primers can use to isolate gene fragment that encodes inulin hydrolysis enzyme from the Bacillus genus.