By means of a zymography (tests of a radial enzyme diffusion in agarose gel and a substrate electrophoresis in polyacrylamide gel) in the preparations of an outer membrane received from strains of a cholera vibrio of O1 and O139 serogroups of different origin and the epidemic importance a range and activity of a certain set of the hydrolytic enzymes which are specific to superficial structures of a bacterial cell have studied. For this purpose as substrate commercial preparations of gelatin, α-lecithin and the protamine sulfate (it is specific to the omptin proteins having enzymatic activity) which were impregnated in agarose and polyacrylamide gels according to copy-books of the standard protocol of the zymographic analysis have been taken. As a result of the conducted researches the hydrolases which are marker for outer membranes of gram negative bacteria – proteases (gelatinases) and lecithinases are found, reliable interstrains differences in degree of their production between toxigenic and nontoxigenic strains are revealed. It is established that the majority of the preparations of an outer membrane taken in a research has protease and lecithinase activity. It is shown that concerning gelatin and the protamin sulfate the preparations of outer membranes received from toxigenic strains of a cholera vibrio while preparations from nontoxigenic strains proteolytic are less active differ in high proteolytic activity. Electrophoretically existence lecithinases in preparations of outer membranes is revealed, however it is established that the lecithinase activity and a spectrum of lecithinases are less than preparations of outer membranes from toxigenic strains, than at nontoxigenic strains. Thus, zymographically the found differences in activity and composition of enzymes allow to carry out further screening of enzymatically active strains of a cholera vibrio for their more detailed studying and designing on their basis as one of components of the acellular vaccine against cholera.

Kozlov Stanislav Nikolaevich, Research Scientist, Irkutsk Anti-plague Research Institute of Siberia and Far East of Rospotrebnadzor, 78, Trilisser st., 664047, Russian Federation, tel.: (3952) 22–01–38, e-mail: ejimei@mail.ru 

Markov Evgeniy Yurievich, Doctor of Sciences (Biology), Senior Research Scientist, Head of department, Irkutsk Anti-plague Research Institute of Siberia and Far East of Rospotrebnadzor, 78, Trilisser st., Irkutsk, 664047, Russian Federation, tel.: (3952) 22–01–38, e-mail: markov_evgenii@mail.ru 

Urbanovich Lyudmila Yakovlevna, Doctor of Sciences (Medicine), Senior Research Scientist, Irkutsk Anti-plague Research Institute of Siberia and Far East of Rospotrebnadzor, 78, Trilisser st., Irkutsk, 664047, Russian Federation, tel.: (3952) 22–01–38 

Nikolaev Valerii Borisovich, Candidate of Sciences (Medicine), Senior Research Scientist, Irkutsk Anti-plague Research Institute of Siberia and Far East of Rospotrebnadzor, 78, Trilisser st., Irkutsk, 664047, Russian Federation, tel.: (3952) 22–01–38, e-mail: balera58.58@mail.ru

Kozlov S.N., Markov E.Yu., Urbanovich L.Ya., Nikolaev V.B. Hydrolase Activity of Vibrio cholerae O1 and O139 Outer Membranes. The Bulletin of Irkutsk State University. Series Biology. Ecology, 2018, vol. 24, pp. 110-117. https://doi.org/10.26516/2073-3372.2018.24.110 (in Russian)

Adamov A.K. Metabolicheskii immunitet k kholere [Metabolic Immunity to Cholera]. Saratov, Saratov St. Univ. Publ., 1982, 176 p.

Markov E.Yu., Urbanovich L.Ya., Golubinskii E.P., Chernov A.B. Naruzhnye membrany kholernogo vibriona kak potentsial'nyi komponent khimicheskoi vaktsiny [Outer Membranes of Vibrio cholerae as Potential Component of Chemical Vaccine]. Zhurn. mikrobiologii [Journal of Microbiology], 1995, no 2, pp. 86-89. (in Russian).

Markov E.Yu., Urbanovich L.Ya., Golubinskii E.P., Karetnikova E.S., Ivanova T.A., Nikolaev V.B., Zavezenov N.P., Subycheva E.N. Poluchenie vysokoimmunogennogo preparata naruzhnykh membran Vibrio cholerae eltor [Production of High Immunogenic Preparation of Vibrio cholerae eltor Outer Membranes]. Zhurn. infektsionnoi patologii [Journal of Infection Pathology], 1998, vol. 5 no. 4, pp. 42-48. (in Russian).

Urbanovich L.Ya., Markov E.Yu., Golubinskii E.P., Kolesnik R.S., Sappo S.G., Ganin V.S., Karetnikova E.S., Ivanova T.A. Kharakteristika immunobiologicheskikh svoistv antigennogo preparata naruzhnykh membran kholernogo vibriona [Description of Immunobiological Properties of Antigenic Preparation of Vibrio cholerae Outer Membranes]. Biotekhnologiya [Biotechnology], 1996, no. 6, pp. 27-33. (in Russian).

Potempa J., Pike R.N. Corruption of innate immunity by bacterial proteases. J. Innate Immun., 2009, vol. 1 no. 12, pp. 70-87. https://doi.org/10.1159/000181144

Nelson D.C., Carbe J., Collin M. Cysteine proteinase SpeB from Streptococcus pyogenes – a potent modifier of immunologically important host and bacterial proteins. Biochem., 2011, vol. 392 no 12, pp. 1077–1088. https://doi.org/10.1515/BC.2011.208

Heussen C., Dowdle E.B. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal. Biochem., 1980, vol. 102 no. 1, pp. 196-202. https://doi.org/10.1016/0003-2697(80)90338-3

Sperandio V., Girón J.A., Silveira W.D., Kaper J.B. The OmpU outer membrane protein, a potential adherence factor of Vibrio cholerae. Infect. Immun., 1995, vol.63 no. 11, pp.4433-4438.