Proteolitic Activity of Marine Strain Bacillus sp. 051
Keywords:Bacillus sp. 051, elastolytic activity, pH optimum, thermal optimum, substrate specificity
The main interest in the study of marine microorganisms is due to their ability to produce a wide range of unique enzymes, including peptidases with different specificities. In recent years, interest has increased in peptidases that are able to cleave elastin as a specific substrate. Streptomyces fradiae and Bacillus thermoproteolyticus elastases are among the most potent elastolytic proteinases discovered to date because they are 4-8-fold more effective than pancreatic elastases. The disadvantages of these producers include the fact that most of them are pathogenic for humans, and the elastase enzyme secreted from them is directly involved in the initiation of the pathogenetic process. All this significantly limits the scope of their practical application. Therefore, the search for new, more effective, safe for humans’ producers continues to be an urgent question, taking into account the fact that there are no highly active elastase producers in Ukraine. Previously we found elastase activity in only 4 of the 10 studied isolates of bacteria from the Black Sea. Since among them, the elastase activity of the Bacillus sp. 051 was the highest, the aim of this work was to study the physicochemical properties and substrate specificity of the enzyme. Methods. We used methods of determining proteolytic (caseinolytic, elastolytic, fibrinolytic, fibrinogenolytic) activity. Protein concentration was determined by the Lowry method. The study of the effect of temperature on the enzymatic activity was carried out in the range from 4 to 70 °C and pH values from 2.0 to 12.0, created using 0.01 M phosphate-citrate buffer. Results. It has been shown that the growing temperature of 12°C is the most optimal for biosynthesis of enzyme by the culture of Bacillus sp. 051. The complex enzyme preparation capable of hydrolyzing elastin, casein and fibrinogen. The enzyme showed maximum activity in relation to elastin (3.65 U/mg). The optimum pH of the enzyme action is 8.0, the thermal optimum is 40°C. The rate of casein hydrolysis compared to elastin was 2.7 times lower and amounted to 1.35 U/mg. The complex enzyme preparation also hydrolyzed fibrinogen (1.16 U/mg). Conclusions. According to its physicochemical and catalytic properties, the representative of the Black Sea, Bacillus sp. 051 is promising for further research as an enzyme producer with elastolytic activity.
Chandrasekaran M., Rajeev Kumar S. Marine Microbial Enzymes Encyclopedia of Life Support Systems (EOLSS) Biotechnology– Vol. IX
Varbanets LD, Avdeeva LV, Borzova NV Matseliukh EV, Gudzenko OV, Kiprianova EA. The Black Sea bacteria-producers of hydrolytic enzymes. Mikrobiol Z. 2011; 73(5): 9–15. Ukrainian.
Gudzenko OV, Avdiyuk KV, Borzova NV, Ivanytsia VО, Varbanets LD. Keratinase, Caseinolitic, Cellulase and β-Mananase Activities of Bacteria Isolated from the Black Sea. Microbiological journal. 2022 (4). P. 3—8. DOI: https://doi.org/10.15407/microbiolj84.04.003
Gudzenko OV., Ivanytsia VО., Varbanets LD. Bacteria of the Black Sea are Producers of α-L-Rhamnosidase. Microbiological journal. 2022 (6). P. 10—15. DOI: https://doi.org/10.15407/microbiolj84.06.010
Varbanets LD., Matseliukh EV. Peptidases of microorganisms and methods of their investigations. Kyiv, Naukova Dumka, 2014, 323 p.
Matseliukh OV, Nidialkova NA, Varbanets LD. [Microbial elastolytic enzyme] Biotechnology Z. 2010; 3(4): 20–28. Ukrainian.
Gudzenko OV., Ivanytsia VО., Varbanets LD. Bacteria of the Black Sea are Producers of Proteolytic Enzymes. Microbiological journal. 2022 (3). P. 3—8. DOI: https://doi.org/10.15407/microbiolj84.03.003
AlShaikh-Mubarak GA, Kotb E, Alabdalall AH, Aldayel MF. A survey of elastase-producing bacteria and characteristics of the most potent producer, Priestia megaterium gasm32. PLoS One. 2023 Mar 13;18(3):e0282963. DOI: https://doi.org/10.1371/journal.pone.0282963
Komori Y, Nonogaki T, Nikai T. Hemorrhagic activity and muscle damaging effect of Pseudomonas aeruginosa metalloproteinase (elastase). Toxicon. 2001;39(9):1327–32. DOI: https://doi.org/10.1016/S0041-0101(01)00084-8
Teufel P, Gotz F. Characterization of an extracellular metalloprotease with elastase activity from Staphylococcus epidermidis. J Bacteriol. 1993;175(13):4218–24. DOI: https://doi.org/10.1128/jb.175.13.4218-4224.1993
Chen Q, Guoqing H, Jinling W. Acid shock of elastase-producing Bacillus licheniformis ZJUEL31410 and its elastase characterization evaluation. J Food Eng. 2007;80(2):490–6. DOI: https://doi.org/10.1016/j.jfoodeng.2006.05.031
Abd EI-Aziz AB, Hassan AA. Optimization of microbial elastase production. J. Radiat. Res. Appl. Sci. 2010;3(4):1237–57.
Lei Y, Zhao P, Li C, Zhao H, Shan Z, Wu Q. Isolation, identification and characterization of a novel elastase from Chryseobacterium indologenes. Appl Biol Chem. 2018;61(3):365–72. DOI: https://doi.org/10.1007/s13765-018-0364-6
Morihara K, Tsuzuki H. Elastolytic properties of various proteinases from microbial origin. Arch Biochem Biophys. 1967;120(1):68–78. doi: 10.1016/0003-9861(67)90599-1 DOI: https://doi.org/10.1016/0003-9861(67)90599-1
Anson ML. The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin. J Gen Physiol. 1938 Sep 20; 22(1):79-89. DOI: https://doi.org/10.1085/jgp.22.1.79
Trombridg GO, Moon HD. Purification of human elastase. Proc Soc Exp Biol Med1972, 141(3):928‒931. DOI: https://doi.org/10.3181/00379727-141-36903
Nidialkova NA, Matseliukh OV , Varbanets LD. Physico-chemical properties of Bacillus thuringiensis IMV B-7324 fibrinolytic peptidase. Mikrobiol Z. 2013; 75(4):3–7. Ukrainian.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with folinphenol reagent. J Biol Chem. 1951; 193(2):265‒275. DOI: https://doi.org/10.1016/S0021-9258(19)52451-6
Lakyn HF. [Byometryia]. Moscow: Vysshaya Shkola; 1990. russian
Tomczak M., Godfrey, JS. Regional Oceanography: An Introduction. Pergamon Press, Oxford. 1994.
Varbanets L.D., Gudzenko O.V., Ivanytsia V.A. Marine Actinobacteria – Producers of Enzymes with α-L-Rhamnosidase. Mikrobiol. Z. 2020; 82(5):3-10. DOI: https://doi.org/10.15407/microbiolj82.05.003
Varbanets LD, Berezkina AE, Avdiuk KV, Gudzenko OV. Keratinolytic and α-l-rhamnosidase activity of bacterial isolates, isolated from gastropod mollusks Nacella concinna (Nacellidae) − residents of Antarctic. Mikrobiol Z. 2020; 82(1):13–21. Ukrainian. DOI: https://doi.org/10.15407/microbiolj82.01.013
Fernandes P. Marine enzymes in food industry. Marine Biot. 2014; 46 (1):1–18. DOI: https://doi.org/10.3389/fmars.2014.00046
How to Cite
Copyright (c) 2023 Mikrobiolohichnyi Zhurnal
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.