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Alkaline serine protease from the new halotolerant alkaliphilic Salipaludibacillus agaradhaerens strain AK-R: purification and properties

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Abstract

Herein, we report the purification and characterization of an alkaline protease from the alkaliphilic Salipaludibacillus agaradhaerens (formerly Bacillus agaradhaerens) strain AK-R, which was previously isolated from Egyptian soda lakes. The purification procedures resulted in enzyme purification up to 13.3-fold, with a recovery yield of 16.3% and a specific activity of 3488 U/mg protein. AK-R protease was a monomeric protein with an estimated molecular weight of 33.0 kDa. The optimum pH and temperature for AK-R protease were pH 10 and 60 °C, respectively. The enzyme thermostability was significantly enhanced in the presence of CaCl2 by approximately 1.3-fold. Moreover, under optimal conditions, the Km and Vmax values of the enzyme were 2.63 mg/ml and 4166.7 U/mg, respectively. PMSF caused complete inhibition of the enzyme activity, suggesting that AK-R belongs to the serine protease family. In addition, the enzyme was completely inhibited by EDTA, revealing the requirement of metal ions for AK-R protease activity; hence, it can be classified as a metalloprotease. AK-R protease is a mostly thiol-independent enzyme, since thiol reductants such as β-mercaptoethanol and dithiothreitol had no effect on the enzyme activity. AK-R protease exhibited high stability in several organic solvents, including butanol, amyl alcohol, dimethyl ether, toluene, diethyl ether and methanol. Moreover, AK-R protease showed significant stability to a variety of surfactants and commercial detergents. The features and properties of AK-R alkaline protease are favourable and suggest its potential applications in various industries, particularly in the laundry detergent industry.

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Funding

This project was supported by King Saud University, Deanship of Scientific Research, College of Science Research Center.

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Authors and Affiliations

  1. Department of Chemistry of Natural and Microbial Products, Pharmaceutical and Drug Industries Research Division, National Research Centre, El-Buhouth St, Dokki, Cairo, 12311, Egypt

    Abdelnasser S. S. Ibrahim & Dina Abdel Fattah Maany

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

    Abdelnasser S. S. Ibrahim, Yahya B. Elbadawi, Mohamed A. El-Tayeb & Khalid S. Al-maary

  3. Department of Biochemistry, College of Science, King Saud University, Riyadh, 11451, Saudi Arabia

    Shebl Salah S. Ibrahim

  4. Tropical Medicine Research Institute, PO Box 1304, Khartoum, Sudan

    Atif A. Elagib

  5. National Centre for Research, Khartoum, Sudan

    Atif A. Elagib

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  1. Abdelnasser S. S. Ibrahim
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  2. Yahya B. Elbadawi
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  3. Mohamed A. El-Tayeb
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  4. Khalid S. Al-maary
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  5. Dina Abdel Fattah Maany
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  6. Shebl Salah S. Ibrahim
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Correspondence to Abdelnasser S. S. Ibrahim.

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Ibrahim, A.S.S., Elbadawi, Y.B., El-Tayeb, M.A. et al. Alkaline serine protease from the new halotolerant alkaliphilic Salipaludibacillus agaradhaerens strain AK-R: purification and properties. 3 Biotech 9, 391 (2019). https://doi.org/10.1007/s13205-019-1928-9

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