Optimization of alkaline protease production from a locally isolated Bacillus sp. ZR-5: Potential application as a detergent additive

Document Type: Research Paper


1 Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

2 Chemical Engineering Department, School of Chemical and Petroleum Engineering, Shiraz University, Shiraz 71345, Iran

3 Department of Biochemistry, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

4 Department of Biology, School of Basic Sciences, University of Isfahan, Isfahan, Iran


The aim of this study was to optimize protease production using a novel Bacillus sp. ZR-5 strain
isolated from the soil, and evaluate its application in detergent industry. The proteolytic activity
of the strain was demonstrated using gelatin hydrolysis screening test. Protease production
optimization was carried out using a two-step approach: a conventional method in order to
identify the best carbon and nitrogen sources followed by the application of response surface
methodology (RSM) to optimize the factors, which include temperature, pH and incubation
time. Glucose or fructose (5 g/L), wheat bran (5 g/L), temperatures of 25 and 55°C, pH 10.0 and
an approximate incubation time of 44 h, were determined as the optimal conditions according to
optimization processes. Validation tests were carried out under these conditions and the results
were in good agreement with RSM predicted data. The in-gel activity (zymogram) test showed
two hydrolytic zones with 66.2 and 36.5 kDa molecular weight on the casein containing
polyacrylamide gel. The high compatibility in the presence of detergent powder and washing
performance test suggested that the crude enzyme could be an appropriate choice as a detergent
additive in detergent industries.


Main Subjects

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