Screening of Protease Producers Among Representatives of the Genus Bacillus Isolated from the Coastal Zone of the Kinburn Split

Authors

  • O.V. Gudzenko Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, 154 Academika Zabolotnoho Str., Kyiv, 03143, Ukraine
  • L.D. Varbanets Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, 154 Academika Zabolotnoho Str., Kyiv, 03143, Ukraine

DOI:

https://doi.org/10.15407/microbiolj86.02.003

Keywords:

Bacillus strains, Kinburn split, caseinolytic, elastolytic, fibrinolytic, fibrinogenolytic, and collagenase activities

Abstract

Protease-producing representatives of Bacillus have been isolated from various environments such as the rhizosphere, soil, sewage, food, waste, and saline environments (sea water, marine sediments, hypersaline lakes, salty food, soda lakes, etc.). However, today, little is known about the characteristics of proteases in representatives of the genus Bacillus, isolated from the coastal and aquatic environments. In this regard, the purpose of this work was to screen for the presence of different types of proteolytic activity in Bacillus cultures isolated from the coastal zone of the Kinburn split. Methods. The objects of the study were 15 cultures (L1-L15), isolated from the dry grass of the coastal zone of the Kinburn spit (Mykolaiv region). The cultures were grown under conditions of deep cultivation at 28 °С, with a mixing speed of the nutrient medium of 230 rpm for 2 days. Methods of determining proteolytic (caseinolytic, elastolytic, fibrinolytic, fibrinogenolytic, and collagenase) activity in the culture liquid supernatant were used. Results. Study of the spectrum of proteolytic activities of 15 strains of Bacillus sp. showed that L9, L1, L2 and L4 exhibited higher levels of proteolytic activity compared to the other strains tested. Bacillus sp. L9 and L2 showed the highest elastase activity (35.80 and 33.80 U/mL, respectively), L1, L2, and L4 − fibrinogenolytic (12.50, 11.20 and 10.33 U/mL, respectively), and L1 and L4 − fibrinolytic (13.00 and 10.90 U/mL, respectively) activity. Conclusions. According to its catalytic properties, a number of representatives of Bacillus sp. isolated from the dry grass of the coastal zone of the Kinburn split may be promising for further research as producers of enzymes with elastotic, fibrinolytic, and fibrinogenolytic activities.

Downloads

Download data is not yet available.

References

Anson, M. L. (1938). The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin. Journal of General Physiology, 22(1), 79–89. https://doi.org/10.1085/jgp.22.1.79

Barzkar, N., Jahromi, S. T., & Vianello, F. (2022). Marine Microbial Fibrinolytic Enzymes: An Overview of Source, Production, Biochemical Properties and Thrombolytic Activity. Marine Drugs, 20(1), 46. https://doi.org/10.3390/md20010046

Contesini, F. J., Melo, R. R. de, & Sato, H. H. (2017). An overview of Bacillus proteases: from production to application. Critical Reviews in Biotechnology, 38(3), 321–334. https://doi.org/10.1080/07388551.2017.1354354

Gudzenko, O. V., Ivanytsia, V. О., & Varbanets, L. D. (2022). Bacteria of the Black Sea Are Producers of Proteolytic Enzymes. Mikrobiolohichnyi Zhurnal, 84(3), 3–8. https://doi.org/10.15407/microbiolj84.03.003

Pernille Lind, Ulrik Hintze, Mogens. (2001). Thrombolytic Therapy Preserves Vagal Activity Early after Acute Myocardial Infarction. Scandinavian Cardiovascular Journal, 35(2), 92–95. https://doi.org/10.1080/140174301750164718

Lowry, Oliver H., Rosebrough, Nira J., Farr, A. L., & Randall, Rose J. (1951). Protein Measurement with the Folin Phenol Reagent. Journal of Biological Chemistry, 193(1), 265–275. https://doi.org/10.1016/s0021-9258(19)52451-6

Macedo, A. J., da Silva, W. O., Gava, R., Driemeier, D., Henriques, J. A., & Termignoni, C. (2005). Novel keratinase from Bacillus subtilis S14 exhibiting remarkable dehairing capabilities. Applied and environmental microbiology, 71(1), 594–596. https://doi.org/10.1128/AEM.71.1.594-596.2005

Moore, S., & Stein, W. H. (1948). Photometric Ninhydrin Method for Use in the Chromatography of Amino Acids. Journal of Biological Chemistry, 176(1), 367–388. https://doi.org/10.1016/s0021-9258(18)51034-6

Nidialkova, N. А., Varbanets, L. D., & Chernyshenko, V. O. (2016). Isolation and purification of Bacillus thuringiensis var. israelensis IMV B-7465 peptidase with specificity toward elastin and collagen. The Ukrainian Biochemical Journal, 88(3), 18–28. https://doi.org/10.15407/ubj88.03.018

Razzaq, A., Shamsi, S., Ali, A., Ali, Q., Sajjad, M., Malik, A., & Ashraf, M. (2019). Microbial Proteases Applications. Frontiers in Bioengineering and Biotechnology, 7. https://doi.org/10.3389/fbioe.2019.00110

Saggu, S. K., & Mishra, P. C. (2017). Characterization of thermostable alkaline proteases from Bacillus infantis SKS1 isolated from garden soil. PLOS ONE, 12(11), e0188724. https://doi.org/10.1371/journal.pone.0188724

Solanki, P., Putatunda, C., Kumar, A., Bhatia, R., & Walia, A. (2021). Microbial proteases: ubiquitous enzymes with innumerable uses. 3 Biotech, 11(10). https://doi.org/10.1007/s13205-021-02928-z

Sumi, H., Yanagisawa, Y., Yatagai, C., & Saito, J. (2004). Natto Bacillus as an Oral Fibrinolytic Agent: Nattokinase Activity and the Ingestion Effect of Bacillus subtilis natto. Food Science and Technology Research, 10(1), 17–20. https://doi.org/10.3136/fstr.10.17

Shuba M. Ya., Fundamentals of Molecular Physiology of Ion Channels, Naukova Dumka, Kyiv (2010), 448 p. (2011). Neurophysiology, 43(1), 75–76. https://doi.org/10.1007/s11062-011-9188-2

Trowbridge, J. O., & Moon, H. D. (1972). Purification of Human Elastase. Experimental Biology and Medicine, 141(3), 928–931. https://doi.org/10.3181/00379727-141-36903

Varbanets, L. D., Matseliuk, O. V., Gudzenko, O. V., Nidialkova, N. A., Zelena, P. P., Yumina, Yu. V., Stepura, L. G., Schepelevich, V. V., & Voychuk, S. I. (2016). Screening of α-L-Rhamnosidases and Peptidases among Actinobacterium and Bacilli. Mikrobiolohichnyi Zhurnal, 78(3), 26–35. https://doi.org/10.15407/microbiolj78.03.026

Yao, Z., Kim, J. A., & Kim, J. H. (2017). Properties of a fibrinolytic enzyme secreted by Bacillus subtilis JS2 isolated from saeu (small shrimp) jeotgal. Food Science and Biotechnology, 27(3), 765–772. https://doi.org/10.1007/s10068-017-0299-4

Yilmaz, M., Soran, H., & Beyatli, Y. (2006). Antimicrobial activities of some Bacillus spp. strains isolated from the soil. Microbiological Research, 161(2), 127–131. https://doi.org/10.1016/j.micres.2005.07.001

Downloads

Published

2024-04-28

How to Cite

Gudzenko, O., & Varbanets, L. (2024). Screening of Protease Producers Among Representatives of the Genus Bacillus Isolated from the Coastal Zone of the Kinburn Split. Mikrobiolohichnyi Zhurnal, 86(2), 3-9. https://doi.org/10.15407/microbiolj86.02.003

Received

2023-09-05

Accepted

2023-10-20

Published

2024-04-28