Screening,selection and development of Bacillus subtilis apr-IBL04 for hyper production of macromolecule alkaline protease |
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Authors: | Tahira Shafique Javeria Shafique Sheikh Zahid Mohsin Kazi Osamah Alnemer Ajaz Ahmad |
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Institution: | 1. IBL4-Department of Biochemistry, University of Agriculture Faisalabad, Pakistan;2. PBL-Department of Bioinformatics and Biotechnology, Govt. College University, Faisalabad, Pakistan;3. Division of Biochemistry, Faculty of Basic Sciences, SKUAST Jammu, J&K 180009, India;4. Department of Pharmaceutics, College of Pharmacy, King Saud University, Saudi Arabia;5. Department of Clinical Pharmacy, College of Pharmacy, King Saud University, Saudi Arabia |
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Abstract: | Bacillus subtilis microbe is commonly found in soil and produces proteases on nitrogen and carbon-containing sources and increases the fertility rate by degrading nitrogenous organic materials. The present study was aimed to develop hyper producing mutant strain of B. subtilis for the production of proteases, to improve the process variables by the response surface methodology (RSM) under central composite design (CCD) and the production of protease by the particular mutant strain in a liquid state fermentation media. The mutation of the strain was carried out using ethidium bromide. Pure B. subtilis strain was collected and screened for hyper-production of protease. The production of protease by mutant B. subtilis strain was optimized by varying temperature, inoculum size, pH and incubation time under liquid state fermentation. The CCD model were found to be reliable with r2 of 0.999. The maximum enzyme activity of B. subtilis IBL-04 mutant with 3 mL/100 mL inoculum size, 72 h fermentation time, pH 8, and 45 °C temperature was developed with enzyme activity 631.09 U/mL, indicates 1–7-fold increase in enzyme activity than the parent strain having 82.32 U/mL activity. These characteristics render its potential use in industries for pharmaceutical and dairy formulation. |
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Keywords: | Alkaline protease Ethidium bromide RSM Enzyme activity |
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