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Characterization of extracellular protease from Bacillus licheniformis

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Research Paper 01/10/2017
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Characterization of extracellular protease from Bacillus licheniformis

Asifa Afzal, Tawaf Ali Shah, Mahjabeen Saleem, Romana Tabassum
Int. J. Biosci.11( 4), 228-236, October 2017.
Certificate: IJB 2017 [Generate Certificate]
Key: Bacillus licheniformisCloneProteaseThermostable

Abstract

Extracellular proteases have a fundamental position with respect to their physiological roles as well as their commercial applications. Bacillus licheniformis RT7P1, was evaluated in this study for production of extracellular protease activity. The culture was maintained on 1% w/v casein plates. In this study, the strain was found to produce maximum enzyme at pH 7 and temperature 37°C after 72 h. The optimum assay pH and the temperature was 10 and 50°C, respectively. The enzyme was stable between pH 9-11and thermal stability data showed that enzyme was stable at 100°C. For cloning of protease gene from Bacillus licheniformis, primers were designed to pick their full-length sequences from the genomic DNA obtained from different Bacillus species. Genomic DNA was isolated from Bacillus licheniformis strain RT7P1 and then the protease gene was amplified from it by using RT7P1 specific primers. This amplified product (1725bp) was then cloned in PTZ57R/T vector. The clone was confirmed by restriction analysis with EcoR1 and BamH1, which showed two fragments of (2886 bp) and (1725 bp), which showed that insert was cloned in the right orientation. The study indicates that Bacillus licheniformis RT7P1 is a good source of commercial thermostable alkaline extracellular protease.

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