Isolation and protease enzyme activity survey in Bacillus isolates collected from Mahalat warm water springs

Protease enzymes are the most commonly used industrial enzymes which have numerous applications in biotechnology. These enzymes have allocated about%60 of the world market industrial enzymes to themselves. Microbes are considered as one of the main sources for producing Protease due to their rapid growth, cultivation ease, and genetic manipulation for optimizing enzymes production. The present study aims to isolate the thermophilic species of Protease-producing Bacillus from Mahallat hot springs in Markazi province and to evaluate the capability for producing Protease enzyme in these species.The initial isolation and identification process was carried out using biochemical tests along with microorganisms with Proteolytic enzyme activity from the Skim milk agar culture. The presence of Protease gene in bacteria species was evaluated using PCR cloning and to identify bacterial species 16 SrRNA gene was amplified by PCR and sequenced. Protease activity was assessed using Lowry test. Finally, the best Bacillus species was detected in terms of Protease activity. 9 bacteria species were selected and isolated as the superior Protease-producing enzyme species using biochemical tests. The analysis of Protease gene and 16 SrRNA gene sequencing indicated that 7 bacteria species have the most proximity to Bacillus species. In enzyme activity assessment, three bacteria species including BN2, DA2, and DA3 demonstrated the highest activity at 47˚C, pH=8, respectively.Due to Protease enzyme various applications in industry, it seems that the use of local strains can help large scale production of Protease enzyme and our country’s self-sufficiency to import this product.