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Isolation, Purification, and Characterization of Protease from a Local Bacillus Strain Adapted to Extreme Temperatures in Southern Iraq

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Title Isolation, Purification, and Characterization of Protease from a Local Bacillus Strain Adapted to Extreme Temperatures in Southern Iraq
 
Creator Alzobaidy, Hyder Najy
Hasan, Ghasan M
Al-Magsoosi, Suhad K R
Awda, Jasim M
 
Subject Bacillus subtilis
Climate change
Enzymes production
Submerged fermentation
Thermostability
 
Description 741-747
Bacteria, with their amenability to genetic manipulation and cultivation, represent a favored source of protease enzymes.
This study aims to isolate and purify protease from a local Bacillus strain in southern Iraq, known for its resilience to
extreme heat. In August 2022, soil samples were collected from Wasit Province, Iraq, and plated on a casein-hydrolyzed
medium. Bacillus strains were selected based on microscopic and colony characteristics, with the most active isolate
identified as Bacillus subtilis through morphological assessment and 16S rRNA sequencing. Protease production occurred
through submerged fermentation, yielding a crude enzyme with enzymatic and specific activities of 241.3 U/mL and 613.3
U/mg, respectively. Partial purification with chilled acetone resulted in an enzyme with enzymatic and specific activities of
935.0 U/mL and 745.0 U/mg, respectively. Further DEAE chromatographic purification showed a single peak of protease
activity with increasing sodium chloride ionic strength. The enzyme's specific activity reached 2491.8 U/mg, with a
recovery rate of nearly 90%. Subsequent gel column filtration via 1.5 × 60 cm Sephadex G-100 displayed a single peak of
protease activity and increased specific activity (4264.10 U/mg) with a 73% recovery rate. Molecular weight determination
using Sephadex G-100 column indicated a size of 29.3 kDa for the B. subtilis protease. Regarding thermal stability, the
enzyme demonstrated initial stability at 60°C, but prolonged exposure reduced activity. In contrast, exposure to 70°C and
80°C resulted in rapid declines in enzymatic activity. This study highlights the isolation and purification of a robust protease
enzyme from a local Bacillus strain, underscoring its potential significance in various industrial applications.
 
Date 2024-07-05T11:29:16Z
2024-07-05T11:29:16Z
2024-07
 
Type Article
 
Identifier 0022-4456 (Print); 0975-1084 (Online)
http://nopr.niscpr.res.in/handle/123456789/64214
https://doi.org/10.56042/jsir.v83i7.6085
 
Language en
 
Publisher NIScPR-CSIR,India
 
Source JSIR Vol.83(7) [July 2024]