The production of alkaline protease by Bacillus tequilensis FJSH2 isolated from Jiroft’s slaughterhouse wastes

Subject Areas : Microbial Biotechnology

Arasto Badoei-dalfard ¹ , Parvin Amiri ² , Narjes Ramezanipour ³ , Zahra Karami ⁴ , Batool Ghanbari ⁵

1 - Assistant Professors, Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.
2 - M.Sc., University of Payam-noor, Tehran, Iran.
3 - M.Sc., Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.
4 - Assistant Professors, Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.
5 - Lecturer, University of Payam-noor, Jiroft, Kerman, Iran.

Received: 2014-07-06 Accepted : 2015-10-01 Published : 2015-05-01

Keywords: Screening, Alkaline protease, Wastes, Bacillus tequilensis,

Abstract :

Background & Objectives: Proteases are the most applied industrial enzymes usable in several biotechnological industries. These enzymes include approximately 60% of enzyme marketing worldwide. Alkaline proteases are the most important enzymes which are stable in the presence of alkaline pH, surfactant and oxidizing agents. This study was aimed to isolate and identify the protease producing bacteria from slaughterhouse wastes in Jiroft. Materials & Methods: This cross - sectional study was performed on Jiroft’s slaughterhouse wastes. The protease producing strain was screened on the skim milk agar media. The 16S rRNA genes of the samples were obtained using PCR and sequencing. The protease enzyme was partially purified by ammonium sulphate precipitation, dialysis and ion exchange chromatography. Thereafter, The biochemical features of the enzymes were analysed. Results: The gene sequencing study showed that the isolated strain were 100 % similar to Bacillus tequilensis species. The purified protease showed maximum activity and stability in pH 9 and 40°C. Km and Vmax values were obtained based on Linewewr berg equation 6.77 mg/ml and 64.94 U/ml/min, respectively. Protease stability was improved about 1.8 and 2.4 folds in the presence of cyclohexane and DMSO, respectively. Conclusion: The activity and stability of this enzyme in alkaline pH and in the presence of organic solvents indicates its potential to be used in industries.