Characterization of extracellular protease produced by Bacillus tequilensis ON1
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Bacillus tequilensis
bùn thải
protease sludge

How to Cite

Nguyễn QL, Nguyễn Đức H, Trịnh TPT, Lê TKT, Lê CT. Characterization of extracellular protease produced by Bacillus tequilensis ON1. hueuni-jns [Internet]. 2022Mar.31 [cited 2022Oct.6];131(1A):109-17. Available from:


A bacterial strain with casein hydrolytic activity was isolated from shrimp waste sludge from a culture pond at Phong Dien, Thua Thien Hue. Molecular identification using 16S rRNA nucleotide sequence shows a high similarity to Bacillus tequilensis on the NCBI database. Hence, the isolate was named B. tequilensis ON1. Extracellular protease activity value is 192,42 U·mL–1 after 24 h culture. Zymogram SDS-PAGE electrophoresis shows that the major extracellular protease has a molecular weight of 130 kDa. The enzyme exhibits maximal activity at 50 °C and pH 8. Protease activity increases in the presence of Ca2+ and Mg2+ ions, while Mn2+ inhibits protease activity. The protease activity was more active and stable in hydrophobic organic solvents.
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  1. Fatma Syahirah M, Nazaitulshila R. The Utilization of Pineapples Waste Enzyme for the Improvement of Hydrolysis Solubility in Aquaculture Sludge. Journal of Energy and Safety Technology. 2019;1(2-2):35-41.
  2. Koyama M, Nagao N, Syukri F, Rahim AA, Kamarudin MS, Toda T, et al. Effect of temperature on thermophilic composting of aquaculture sludge: NH3 recovery, nitrogen mass balance, and microbial community dynamics. Bioresource technology. 2018;265:207-13.
  3. Hopkins JS, Sandifer PA, Browdy CL. Sludge management in intensive pond culture of shrimp: Effect of management regime on water quality, sludge characteristics, nitrogen extinction, and shrimp production. Aquacultural Engineering. 1994;13(1):11-30.
  4. Domingues RF, Sanches T, Silva GS, Bueno BE, Ribeiro R, Kamimura ES, et al. Effect of Enzymatic Pre-Treatment On The Anaerobic Digestion Of Milk Fat For Biogas Production. Food Research International. 2015;73:26-30.
  5. Contesini FJ, Melo RR, Sato HH. An overview of Bacillus proteases: from production to application. Critical reviews in biotechnology. 2018;38(3):321-34.
  6. Sambrook J, Maccallum P, Russell D. Molecular Cloning: A Laboratory Manua. 3rd, editor. Cold Spring Harbor Press: NY; 2001.
  7. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution. 2018;35:1547-9
  8. Cupp-Enyard C. Sigma's Non-specific Protease Activity Assay - Casein as a Substrate. Journal of Visualized Experiments. 2008;19:899.
  9. Wang Q, Ji F, Wang J, Jiang B, Li L, An L, et al. Characterization of a salt-activated protease with temperature dependent secretion in Stenotrophomonas maltophilia FF11 isolated from frozen Antarctic krill. Journal of Industrial Microbiology & Biotechnology. 2017;43(6):829-40.
  10. Karray A, Alonazi M, Horchani H, Ben Bacha, Novel AA. Thermostable and Alkaline Protease Produced from Bacillus stearothermophilus Isolated from Olive Oil Mill Sols Suitable to Industrial Biotechnology. Molecules. 2021;26:1-15.
  11. Panta G, Prakasha A, Beraa JVPPS, Panchpuri GVNSD, Kumara A, Panchpuri M, et al. Production, optimization and partial purification of protease from Bacillus subtilis. Journal of Taibah University for Science. 2015;9(1):50-5.
  12. Khan Z, Shafique M, Nawaz HR, Jabeen N, Naz SA. Bacillus tequilensis ZMS-2: A novel source of alkaline protease with antimicrobial, anti-coagulant, fibrinolytic and dehairing potentials Pakistan Journal of Pharmaceutical Sciences. 2019;23(4):1913-8.
  13. Maruthiah T, Immanuel G, Palavesam A. Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2015;87(1):207-16.
  14. Robinson, PK. Enzymes: principles and biotechnological applications. Essays in biochemistry. 2015;59:1-41.
  15. Hammami A, Fakhfakh N, Abdelhedi O, Nasri M, Bayoudh A. Proteolytic and amylolytic enzymes from a newly isolated Bacillus mojavensis SA: Characterization and applications as laundry detergent additive and in leather processing. International Journal of Biological Macromolecules. 2018;108:56-68.
  16. Yilmaz B, Baltaci MO, Sisecioglu M, Adiguzel A. Thermotolerant alkaline protease enzyme from Bacillus licheniformis A10: purification, characterization, effects of surfactants and organic solvents. Journal of Enzyme Inhibition and Medicinal Chemistry. 2015;31(6):1241-7.
  17. Benkiar A, Nadia Z, Badis A, Rebzani F, Soraya B, Rekik H, et al. Biochemical and molecular characterization of a thermo- and detergent-stable alkaline serine keratinolytic protease from Bacillus circulans strain DZ100 for detergent formulations and feather-biodegradation process. International Biodeterioration and Biodegradation. 2013;83:129-38.
  18. Huang S, Pan S, Chen G, Huang S, Zhang Z, Li Y, et al. Biochemical characteristics of a fibrinolytic enzyme purified from a marine bacterium, Bacillus subtilis HQS-3. International Journal of Biological Macromolecules. 2013;62:124-30.
  19. Sharma KM, Kumar R, Panwar S, Kumar A. Microbial alkaline proteases: Optimization of production parameters and their properties. Journal of Genetic Engineering and Biotechnology. 2017; 15(1):115-126.
  20. Razzaq A, Shamsi S, Ali A, Ali Q, Sajjad M, Malik A, Ashraf M. Microbial proteases applications. Frontiers in Bioengineering and Biotechnology. 2019;7:110.
  21. Abd Rahman RNZR, Mahamad S, Salleh AB, Basri M. A new organic solvent tolerant protease from Bacillus pumilus 115b. Journal of Industrial Microbiology & Biotechnology. 2007;34(7):509-17.
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