3 Biotech

, 9:391 | Cite as

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

  • Abdelnasser S. S. IbrahimEmail author
  • Yahya B. Elbadawi
  • Mohamed A. El-Tayeb
  • Khalid S. Al-maary
  • Dina Abdel Fattah Maany
  • Shebl Salah S. Ibrahim
  • Atif A. Elagib
Original Article


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.


Alkaline proteases Salipaludibacillus agaradhaerens Soda lakes Alkaliphilic bacteria 



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

Compliance with ethical standards

Conflict of interest

There is no conflict of interest.

Supplementary material

13205_2019_1928_MOESM1_ESM.docx (57 kb)
Supplementary material 1 (DOCX 56 kb)


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Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  • Abdelnasser S. S. Ibrahim
    • 1
    • 2
    Email author
  • Yahya B. Elbadawi
    • 2
  • Mohamed A. El-Tayeb
    • 2
  • Khalid S. Al-maary
    • 2
  • Dina Abdel Fattah Maany
    • 1
  • Shebl Salah S. Ibrahim
    • 3
  • Atif A. Elagib
    • 4
    • 5
  1. 1.Department of Chemistry of Natural and Microbial Products, Pharmaceutical and Drug Industries Research DivisionNational Research CentreCairoEgypt
  2. 2.Department of Botany and Microbiology, College of ScienceKing Saud UniversityRiyadhSaudi Arabia
  3. 3.Department of Biochemistry, College of ScienceKing Saud UniversityRiyadhSaudi Arabia
  4. 4.Tropical Medicine Research InstituteKhartoumSudan
  5. 5.National Centre for ResearchKhartoumSudan

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