Screening and Production of Peptidase Producing Bacillus Species Isolated from Soil

Abstract

Aims: Proteases are the most deliberate microbial enzyme at mercantile, industrial, health professional, analytic, symptomatic, outgoing interruption and other sectors. Methodology: About 8 isolates were recovered from different soil samples gathered from various fields of Abdul Hakeem District Khanewal of Pakistan. Bacterial identification was performed by Gram’s staining, endospore staining and hang drop motility test. Samples after serial dilution plated on gelatin agar plates which have composition, Nutrient agar 7.5 gram, gelatin 21 gram, NaCl 0.15 gram, Agar,1.5 gram and 300 ml of distilled water. On these plates zone of inhibition appeared after incubation of 48 hours. The enzyme was extracted by ammonium sulphate precipitation technique. Protease enzyme was characterized by Fourier Transform Infrared Spectroscopy. Enzyme producing activity of bacterial isolate was studied by using spectrophotometer at 280 nm. Results: Among these, three isolates (SA1, SA2, SA3) showed good proteolysis activity, two isolates display moderate activity (SA7, SA8) while the rest were inactive. The plates with clear zone of inhibition were selected for quantitative tests of protease and colonies of bacteria were shifted onto new substrate media plates by wire loop for further testing. Protease assay was performed on enzyme solution at pH 8. The spectral peaks were observed in the region of 1000-3500 cm-1 are 3227.03, 1636.55, 1445.50 and 1061.13 cm-1. The C-H group appeared in the region 3227.03 cm-1. Conclusion: These proteases producing Bacillus species has positive impact in biotechnology and can be conveniently considered for further industrial applications.