Применение зимографического и протеомного анализа для идентификации внутриклеточных и экстрацеллюлярных протеаз производственных штаммов Vibrio cholerae

摘要: The aim of this work was to study the composition and functions intracellular extracellular proteases production Vibrio cholerae strains 569B serovar Inaba M41 Ogawa using zymographic proteomic analysis. Materials methods . Samples were obtained from cell lysates by ultrasonication bacterial cells in a 9 M urea solution. protease fraction precipitated culture liquid adding 50 % trichloroacetic acid final concentration 10 incubating on ice. Lyophilized preparations proteinase K proteovibrin enzyme complex used as control proteolytic activity. Proteases detected substrate gel electrophoresis 12.5 polyacrylamide impregnated with 0.1 gelatin, followed identification protein fractions exoproteins both molecular mass spectrometric scanning. Results discussion A comparative V. analysis showed that greatest enzymatic activity sample strain, where five major four minor zones gelatin hydrolysis identified, high-intensity MW 20–23 37–40 kDa also found preparation isolated fluid strain. As result studied strains, 66 enzymes different functional reliably among which 15 had high information content modern provided for possibility identifying qualitative quantitative differences , offers an effective means screening inter-strain spectrum strains.