Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Synthesis, Optimization, and Characterization of Cellulase Enzyme Obtained from Thermotolerant Bacillus subtilis F3: An Insight into Cotton Fabric Polishing Activity

  • Amr Fouda (Botany and Microbiology Department, Faculty of Science, Al-Azhar University) ;
  • Khalid S. Alshallash (Department of Biology, College of Science, Imam Mohammad Ibn Saud Islamic University) ;
  • Hossam M. Atta (Botany and Microbiology Department, Faculty of Science, Al-Azhar University) ;
  • Mamdouh S. El Gamal (Botany and Microbiology Department, Faculty of Science, Al-Azhar University) ;
  • Mohamed M. Bakry (Botany and Microbiology Department, Faculty of Science, Al-Azhar University) ;
  • Abdullah S. Alawam (Department of Biology, College of Science, Imam Mohammad Ibn Saud Islamic University) ;
  • Salem S. Salem (Botany and Microbiology Department, Faculty of Science, Al-Azhar University)
  • Received : 2023.09.14
  • Accepted : 2023.10.04
  • Published : 2024.01.28
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Abstract

The efficacy of 40 bacterial isolates obtained from hot spring water samples to produce cellulase enzymes was investigated. As a result, the strain Bacillus subtilis F3, which was identified using traditional and molecular methods, was selected as the most potent for cellulase production. Optimization was carried out using one-factor-at-a-time (OFAT) and BOX-Behnken Design to detect the best conditions for the highest cellulase activity. This was accomplished after an incubation period of 24 h at 45℃ and pH 8, with an inoculum size of 1% (v/v), 5 g/l of peptone as nitrogen source, and 7.5 g/l of CMC. Moreover, the best concentration of ammonium sulfate for cellulase enzyme precipitation was 60% followed by purification using a dialysis bag and Sephadex G-100 column chromatography to collect the purified enzyme. The purified cellulase enzyme was characterized by 5.39-fold enrichment, with a specific activity of 54.20 U/mg and a molecular weight of 439 kDa. There were 15 amino acids involved in the purified cellulase, with high concentrations of 160 and 100 mg/l for glycine and proline respectively. The highest stability and activity of the purified cellulase was attained at pH 7 and 50℃ in the presence of 150 ppm of CaCl2, NaCl, and ZnO metal ions. Finally, the biopolishing activity of the cellulase enzyme, as indicated by weight loss percentages of the cotton fabric, was dependent on concentration and treatment time. Overall, the thermotolerant B. subtilis F3 strain has the potential to provide highly stable and highly active cellulase enzyme for use in biopolishing of cotton fabrics.

Keywords

Acknowledgement

The authors extend their appreciation to the Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo, Egypt, for the great support in the achievement and publication of this research work. Furthermore, the authors acknowledge the support from Imam Mohammad Ibn Saud Islamic University Riyadh, Saudi Arabia.

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