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Detergent-, solvent- and salt-compatible thermoactive alkaline serine protease from halotolerant alkaliphilic Bacillus sp. NPST-AK15: purification and characterization

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Abstract

Alkaline protease produced by the halotolerant alkaliphilic Bacillus sp. strain NPST-AK15 was purified to homogeneity by the combination of ammonium sulfate precipitation, anion-exchange and gel permeation chromatography. The purified enzyme was a monomeric protein with an estimated molecular weight of 32 kDa. NPST-AK15 protease was highly active and stable over a wide pH range, with a maximal activity at pH 10.5. The enzyme showed optimum activity at 60 °C and was stable at 30–50 °C for at least 1 h. Thermal stability of the purified protease was substantially improved by CaCl2 (1.1- to 6.6-fold). The K m, V max and k cat values for the enzyme were 2.5 mg ml−1, 42.5 µM min−1 mg−1, and 392.46 × 103 min−1, respectively. NPST-AK15 protease activity was strongly inhibited by PMSF, suggesting that the enzyme is a serine protease. The enzyme was highly stable in NaCl up to 20 % (w/v). Moreover, the purified enzyme was stable in several organic solvents such as diethyl ether, benzene, toluene, and chloroform. In addition, it showed high stability and compatibility with a wide range of surfactants and commercial detergents and was slightly activated by hydrogen peroxide. These features of NPST-AK15 protease make this enzyme a promising candidate for application in the laundry and pharmaceutical industries.

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Acknowledgments

This project was funded by the National Plan for Science, Technology and Innovation (MAARIFAH), King Abdulaziz City for Science and Technology, Kingdom of Saudi Arabia, Award Number (12-BIO2899-02).

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Authors and Affiliations

  1. Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, 11451, Kingdom of Saudi Arabia

    Abdelnasser S. S. Ibrahim, Ali A. Al-Salamah, Yahya B. El-Badawi & Mohamed A. El-Tayeb

  2. Department of Chemistry of Natural and Microbial Products, National Research Center, El-Buhouth St., Dokki, Cairo, 12311, Egypt

    Abdelnasser S. S. Ibrahim

  3. Institute of Technical Microbiology, Hamburg University of Technology, 21073, Hamburg, Germany

    Garabed Antranikian

Authors
  1. Abdelnasser S. S. Ibrahim
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  2. Ali A. Al-Salamah
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  3. Yahya B. El-Badawi
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  4. Mohamed A. El-Tayeb
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Correspondence to Abdelnasser S. S. Ibrahim.

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Communicated by M. da Costa.

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Ibrahim, A.S.S., Al-Salamah, A.A., El-Badawi, Y.B. et al. Detergent-, solvent- and salt-compatible thermoactive alkaline serine protease from halotolerant alkaliphilic Bacillus sp. NPST-AK15: purification and characterization. Extremophiles 19, 961–971 (2015). https://doi.org/10.1007/s00792-015-0771-0

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  • DOI: https://doi.org/10.1007/s00792-015-0771-0

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