Tag Archives: elastolytic and fibrinogenolytic activity
Purification and physicochemical properties of Bacillus licheniformis 249 protease with elastolytic and fibrin(ogen)olitic activity
, , , ,
1Danylo Zabolotny Institute of Microbiology and Virology,
National Academy of Sciences of Ukraine, Kyiv;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Odesa I. I. Mechnikov National University, Odesa, Ukraine;
*e-mail: alena.gudzenko81@gmail.com
Received: 22 April 2026; Revised: 27 May 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Proteolytic enzymes with a broad substrate specificity are of considerable interest for potential application in biotechnology and medicine. Proteases with specificity for elastin, fibrinogen and fibrinogen are of particular interest, in particular as antithrombotic agents. Marine microorganisms due to adaptive mechanisms to environment could be a potential producers of enzymes capable of maintaining catalytic activity in a wide range of physicochemical conditions. Objective. The aim of this work was to purify the protease produced by Bacillus licheniformis 249, isolated from the bottom sediments of the Black Sea and to study the physicochemical and catalytic properties as well as substrate specificity of enzyme. Methods. The enzyme was obtained from the supernatant of the bacterial culture liquid by precipitation with ammonium sulfate (90% saturation) with subsequent gel-permeation chromatography on Toyopearl HW-65F and ion-exchange chromatography on Toyopearl DEAE-650M. Results. The purified enzyme had a molecular weight of about 30.0 kDa, maximal activity at 37°C, pH 8,0 and 11,0 and was characterized by high elastolytic, fibrinogenolytic and fibrinolytic activities with a values of 2697.3; 1620 and 1615 U/mg protein, respectively. Electrophoretic analysis of fibrinogen hydrolysis products revealed selective cleavage of the Aα chain, which allows us to attribute the studied enzyme to α-fibrinogenases. Conclusion. The combination of elastolytic and fibrin(ogen)olytic activities, as well as stability in a wide range of physicochemical conditions, indicate the promising potential of this enzyme for use in medicine and various biotechnological processes.
Purification and physico-chemical properties of Bacillus atrophaeus protease with elastolytic and fibrinogenolytic activity
O. V. Gudzenko1*, L. D. Varbanets1, V. O. Chernyshenko2,
Y. M. Stohnii2, A. M. Ostapchuk3, V. O. Ivanytsia3
1Institute of Microbiology and Virology named after D. K. Zabolotny,
National Academy of Sciences of Ukraine, Kyiv;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Odesa I.I. Mechnikov National University, Odesa, Ukraine;
*e-mail: alena.gudzenko81@gmail.com
Received: 18 September 2024; Revised: 21 October 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Microbial proteases, among which proteases capable of cleaving elastin, fibrin, fibrinogen, and collagen, have been a matter of interest to researchers due to their significant biotechnological potential along with low production cost. We previously showed that Bacillus atrophaeus 08 synthesizes an extracellular protease complex that exhibits high elastolytic, fibrinogenolytic, fibrinolytic activity, and minor caseinolytic and collagenase activity. The aim of the work was to isolate and purify the Bacillus atrophaeus 08 protease from the culture liquid supernatant and to study the physicochemical properties and substrate specificity of enzyme preparation. Precipitation with ammonium sulfate of 90% saturation, gel-permeation and ion-exchange chromatography were used in the experiment. According to the data obtained, the yield of the purified enzyme with a molecular weight of about 30 kDa was 6%, its elastase activity increased 30 times (420 U/mg protein), and fibrinogenolytic activity 31.8 times (350 U/mg protein). In addition, it also exhibited fibrinolytic (35.3 U/mg protein), minor caseinolytic activity (1.2 U/mg protein) and no collagenase activity. The optimum of elastin hydrolysis was at 37°C, pH 3.0 and 9.0-10.0, the optimum for fibrinogen hydrolysis was 12°C, pH 4.0. SDS-PAAG electrophoresis showed that the Bβ-chain of fibrinogen was almost not cleaved even after 1 h of incubation with the enzyme, while the Aα-chain disappeared already at the 30th min with the production of fragments with M.W. of about 30-45 kDa. The activity of the studied enzyme preparation towards fibrin was much lower than towards fibrinogen.