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Alkaline Protease Production Using Response Surface Methodology, Characterization and Industrial Exploitation of Alkaline Protease of Bacillus subtilis sp.

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Abstract

Bacterial proteases are considered potential candidates for industrial and biotechnological applications. In the present study, the influence of various parameters on an in-house isolated alkaline protease was investigated through central composite design from a well-known response surface methodology technique. A high level of alkaline protease productivity up to 8.9-folds was achieved from a novel strain of Bacillus subtilis isolated from local Tannery, Lahore Distt. (Pakistan). The purification profile revealed 8.48-fold increase to homogeneity by ammonium sulfate fractionation and Sephadex G-100-based gel filtration chromatography. The purified fraction was identified as a monomeric with apparent molecular weight of 15 kDa on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The catalytic characterization was performed with various substrates using Michaelis–Menten equation. The alkaline protease remained optimally active within a broader pH range of 8–12 having a half-life of 37.8 min at 87 °C, along with a maximum velocity (Vmax) of 200 IU/mL/min and 0.090 mg/mL Michaelis constants. Moreover, a complete de-staining and dehairing were recorded within the short incubation period. In conclusion, the data thus obtained in this study suggest a high potential for enzymatic treatment in an industrial setting.

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Acknowledgements

This article is based on “Kinetic study and industrial application of protease” funded by Higher Education Commission of Pakistan (PM-IPFP/HRD/HEC/2012/3556). The financial support provided by HEC is thankfully acknowledged.

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Authors and Affiliations

  1. Department of Applied Chemistry & Biochemistry, Government College University, Faisalabad, 38000, Pakistan

    Fouzia Hussain, Shagufta Kamal & Saima Rehman

  2. Department of Botany, Government College University, Faisalabad, 38000, Pakistan

    Muhammad Azeem

  3. Department of Chemistry, Islamia University, Bahawalpur, 63100, Pakistan

    Ismat Bibi

  4. Department of Statistics, Government College University, Faisalabad, 38000, Pakistan

    Tanvir Ahmed

  5. School of Engineering and Science, Tecnologico de Monterrey, Campus Monterrey, Ave. Eugenio Garza Sada 2501, CP 64849, Monterrey, NL, Mexico

    Hafiz M. N. Iqbal

Authors
  1. Fouzia Hussain
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  2. Shagufta Kamal
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  3. Saima Rehman
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  4. Muhammad Azeem
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  5. Ismat Bibi
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  6. Tanvir Ahmed
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  7. Hafiz M. N. Iqbal
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Correspondence to Hafiz M. N. Iqbal.

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Hussain, F., Kamal, S., Rehman, S. et al. Alkaline Protease Production Using Response Surface Methodology, Characterization and Industrial Exploitation of Alkaline Protease of Bacillus subtilis sp.. Catal Lett 147, 1204–1213 (2017). https://doi.org/10.1007/s10562-017-2017-5

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  • DOI: https://doi.org/10.1007/s10562-017-2017-5

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