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Biochemical characterization of a halophilic, alkalithermophilic protease from Alkalibacillus sp. NM-Da2

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Abstract

An extracellular, halophilic, alkalithermophilic serine protease from the halo-alkaliphilic Alkalibacillus sp. NM-Da2 was purified to homogeneity by ethanol precipitation and anion-exchange chromatography. The purified protease was a monomeric enzyme with an approximate molecular mass of 35 kDa and exhibited maximal activity at 2.7 M NaCl, pH55 °C 9 and 56 °C. The protease showed great temperature stability, retaining greater than 80 % of initial activity after 2 h incubation at 55 °C. The protease was also extremely pH tolerant, retaining 80 % of initial activity at pH55 °C 10.5 after 30 min incubation. Protease hydrolyzed complex substrates, displaying activity on yeast extract, tryptone, casein, gelatin and peptone. Protease activity was inhibited at casein concentrations greater than 1.2 mg/mL. The enzyme was stable and active in 40 % (v/v) solutions of isopropanol, ethanol and benzene and was stable in the presence of the polysorbate surfactant Tween 80. Activity was stimulated with the oxidizing agent hydrogen peroxide. Inhibition with phenyl methylsulfonylfluoride indicates it is a serine protease. Synthetic saline wastewater treated with the protease showed 50 % protein removal after 5 h. Being halophilic, alkaliphilic and thermophilic, in addition to being resistant to organic solvents, this protease has potential for various applications in biotechnological and pharmaceutical industries.

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Acknowledgments

This work was supported by the US, Egypt Science and Technology Joint Fund in cooperation with the Suez Canal University (Egypt) under Project number 1841 and the University of Georgia (USA) under Project Number NSF-OISE-1132412.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt

    Asmaa R. Abdel-Hamed, Dina M. Abo-Elmatty & Noha M. Mesbah

  2. Department of Microbiology, University of Georgia, Athens, GA, 30602, USA

    Juergen Wiegel

Authors
  1. Asmaa R. Abdel-Hamed
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  2. Dina M. Abo-Elmatty
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  3. Juergen Wiegel
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  4. Noha M. Mesbah
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Correspondence to Noha M. Mesbah.

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Communicated by L. Huang.

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Abdel-Hamed, A.R., Abo-Elmatty, D.M., Wiegel, J. et al. Biochemical characterization of a halophilic, alkalithermophilic protease from Alkalibacillus sp. NM-Da2. Extremophiles 20, 885–894 (2016). https://doi.org/10.1007/s00792-016-0879-x

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  • DOI: https://doi.org/10.1007/s00792-016-0879-x

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