Background: Enzyme technology has become a new and practical method that has been able to solve many problems and improve many industrial processes, including the production of pharmaceuticals and human food. Among industrial enzymes, cellulases occupy second place in the global enzyme market. Betaglucanases are one of the types of cellulases, which account for a major share of this base. Many groups of prokaryotes and unicellular eukaryotes such as fungi produce this enzyme. Pectobacterium carotovorum is a plant pathogen that secretes important extracellular enzymes, one of which is beta-glucanase. Therefore, the isolation of the beta-glucanase gene of this bacterium and its homogenization in the expression vector of Escherichia coli (pET26b; pSL1180) were considered. Materials and Methods: The Pectobacterium carotovorum was cultured, its genomic DNA was extracted and used to replicate the beta-glucanase coding gene.Then this gene was replicated and cut by an enzyme using specific primers with a cutting site. In parallel, a plasmid was extracted and cut with an enzyme. The betaglucanase gene was inserted into the plasmid and the susceptible host cell was transformed using the product of the transfection reaction and cultured on the selected culture medium. After proving the presence of the recombinant construct, this construct was extracted and the host cell was transformed using it, and the expression of the recombinant beta-glucanase was checked using SDS-PAGE.
Result: The result of electrophoresis on agarose gel, Colony-PCR experiments, and enzymatic digestion and sequencing showed that the beta-glucanase gene was successfully amplified and Cloning in pSL1180, but its subcloning was not successful in pET26b. Nucleoid hybridization BLAST results showed the greatest
similarity with Pectobacterium polaris species. The translation of the synonymous base to amino acid and BLAST confirmed the above data. The result of SDS-PAGE showed the cytoplasmic expression of the
