Detection of hydrolases of different subclasses in cell-free fractions of Vibrio cholerae O1 and O139 serogroups using radial enzymatic diffusion in agarose gel

Abstract

Cholera remains an actual infection worldwide, which dictates the need for a comprehensive study of its pathogens and, in particular, their hydrolytic enzymes that interact with the cells of the macroorganism. We determined the presence of hydrolases from different subclasses in preparations of cell-free fractions obtained from 58 strains of Vibrio cholerae O1 and O139 serogroups of different epidemic significance and origins using radial enzyme diffusion in agarose gels with various substrates. The analysis revealed statistically significant differences in enzyme activity depending on the origin and epidemiological significance of the original strain. We found that preparations obtained from non-toxigenic strains had increased activity of proteases and chitinolytic enzymes, while those from toxigenic strains showed high activity of mucinases, lipolytic enzymes, and nucleases. These data can be applied in the microbiological diagnosis of cholera as an additional biochemical characterization of Vibrio cholerae strains, as well as contribute to the understanding of the role of enzymes in the pathogenesis and adaptation of cholera pathogens.