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Purification and biochemical characterization of two detergent-stable serine alkaline proteases from Streptomyces sp. strain AH4

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Abstract

Streptomyces sp. strain AH4 exhibited a high ability to produce two extracellular proteases when cultured on a yeast malt-extract (ISP2)-casein-based medium. Pure proteins were obtained after heat treatment (30 min at 70 °C) and ammonium sulphate fractionation (30–60 %), followed by size exclusion HPLC column. Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis revealed that the purified enzymes (named SAPS-P1 and SAPS-P2) were monomers with molecular masses of 36,417.13 and 21,099.10 Da, respectively. Their identified N-terminal amino acid displayed high homologies with those of Streptomyces proteases. While SAPS-P1 was optimally active at pH 12.0 and 70 °C, SAPS-P2 showed optimum activity at pH 10.0 and 60 °C. Both enzymes were completely stable within a wide range of temperature (45–75 °C) and pH (8.0–11.5). They were noted to be completely inhibited by phenylmethanesulfonyl fluoride and diisopropyl fluorophosphates, which confirmed their belonging to the serine proteases family. Compared to SAPS-P2, SAPS-P1 showed high thermostability and excellent stability towards bleaching, denaturing, and oxidizing agents. Both enzymes displayed marked stability and compatibility with a wide range of commercial laundry detergents and significant catalytic efficiencies compared to Subtilisin Carlsberg and Protease SG-XIV. Overall, the results indicated that SAPS-P1 and SAPS-P2 can be considered as potential promising candidates for future application as bioadditives in detergent formulations.

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Acknowledgments

This work was funded by the Algerian Ministry of Higher Education and Scientific Research and Technology and the Tunisian Ministry of Higher Education and Scientific Research (Project Tuniso-Algerian JAOUADI/BADIS Grant No. TA/04/2012_TNDZ-MicrooZymes_2012-2015). We wish to express our gratitude to Mr. B. Naili and M. Hmidi (LMBEE-CBS), Mrs. N. Masmoudi and Mr. K. Walha (Analysis Unit-CBS) for their technical assistance and to Dr. A. Beji (Max-Planck-Institute of Biochemistry, München, Germany) and Dr. S. Kourdali (CNRDPA) for their productive discussions and valuable help during the preparation of this study. Special thanks are also due to Mr. A. Smaoui and Mrs. H. Ben Salem from the English department at the Sfax Faculty of Science (Sfax, Tunisia) for their constructive language polishing and editing services. The authors would also like to extend their thanks to the anonymous reviewers for their valuable comments and efforts during the revision of the present manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratory of Microbial Systems Biology (LMSB), École Normale Supérieure (ENS) de Kouba, PO Box 92, 16050, Kouba, Algiers, Algeria

    Souraya Boulkour Touioui & Hadjira Boudjella

  2. Laboratory of Natural Products Chemistry and Biomolecules (LNPCB), University of Blida, 1, Road of Soumaâ, PO Box 270, 09000, Blida, Algeria

    Souraya Boulkour Touioui, Samir Bejar & Bassem Jaouadi

  3. Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, 3018, Sfax, Tunisia

    Nadia Zaraî Jaouadi, Mouna Belhoul & Hatem Rekik

  4. National Centre for Research and Development of Fisheries and Aquaculture (CNRDPA), 11, Bd Amirouche, PO Box 67, 42415, Bou Ismaïl, Tipaza, Algeria

    Fatma Zohra Ferradji & Abdelmalek Badis

  5. Laboratory of Natural Substances Chemistry and Biomolecules, University of Blida, 1, Road of Soumaâ, PO Box 270, 09000, Blida, Algeria

    Abdelmalek Badis

Authors
  1. Souraya Boulkour Touioui
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  2. Nadia Zaraî Jaouadi
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  3. Hadjira Boudjella
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  4. Fatma Zohra Ferradji
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  5. Mouna Belhoul
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  6. Hatem Rekik
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  7. Abdelmalek Badis
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  8. Samir Bejar
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  9. Bassem Jaouadi
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Correspondence to Abdelmalek Badis or Bassem Jaouadi.

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Boulkour Touioui, S., Zaraî Jaouadi, N., Boudjella, H. et al. Purification and biochemical characterization of two detergent-stable serine alkaline proteases from Streptomyces sp. strain AH4. World J Microbiol Biotechnol 31, 1079–1092 (2015). https://doi.org/10.1007/s11274-015-1858-6

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  • DOI: https://doi.org/10.1007/s11274-015-1858-6

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