Mikrobiol. Z. 2019; 81(4):3-14.
Extracellular Lectin Produced by Bacillus subtilis Strain IMV B-7014
Depending on the Culture Conditions
Kisten O.G.1, Kovalenko E.O.1, Getman K.I.1, Sashchuk O.V.2, Pidgorsky V.S.1, Tyshchenko L.M.3
1Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
2Odesa Mechnikov National University
2 Dvoryanskaya Str., Odesa, 65082, Ukraine
3National University of Life and Environmental Sciences of Ukraine
15 Heroiv Oborony Str., Kyiv, 03041, Ukraine
The aim. To develop a new nutrient medium for the biosynthesis of extracellular lectin strain Bacillus subtilis IMB B-7014; to study changes in biomass and pH values, accumulation of protein metabolites in the culture fluid (CF) and in its foamed fraction under separate batch cultivation of the R and S morphotypes of the strain in the developed medium. Methods. Microbiological, biochemical, immunological. The selection of the source of inorganic nitrogen (ammonium citrate, ammonium nitrate, ammonium sulfate, glycine, diammonium phosphate, urea) was performed using the base medium of the following composition, g/l: galactose – 10.0; yeast extract – 2.0; K2HPO4 – 0.70; KH2PO4 – 0.30; NaCl – 0.50; MgSO4∙7H2O – 0.50; CaCl2∙6H2O – 0.10; FeSO4∙7H2O – 0.005; CoCl2∙6H2O – 0.005; MnSO4∙4H2O – 0.005; pH 7.0. Lectin activity (LA) of the CF of both morphotypes of the strain evaluated by the level of hemagglutination activity of its supernatant with rabbit erythrocytes. The molecular weight of the protein elements was determined in a denaturing system (SDS-PAGE). Results. The highest values of LA of the CF observed using ammonium sulfate. Differences in the dynamics of changes in optical density and pH of the CF during batch cultivation of the R and S morphotypes of the strain not observed, whereas the LA of the R morphotype CF was higher. Major and sub-major protein components with a molecular weight of about 50 and 73 kDa, respectively, found in the CF and in its foamed fraction of both strain morphotypes. A unique component with a molecular weight of 64.5 kDa was present in the foam. A component with a molecular weight of 84 kDa was present on the electrophoregrams of the CF R and S morphotypes in an insignificant amount, but was absent in the foam fraction. A sub-major component with a molecular weight of 43 kDa of a foam fraction found in the CF S and R morphotypes in trace amounts. The foam fraction characterized by a large presence of protein components due to their transition from the CF. Conclusions. The LA of the CF R morphotype was higher than for the S morphotype. The foam fraction contained more protein components than the CF.
Keywords: Bacillus subtilis, R and S strain morphotype, nitrogen source, extracellular lectin, culture fluid, foam, protein components.
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