ABSTRACT
AIM: Study N-acetyl-ß-D-glucosaminidase (chitobiase) (EC 3.2.1.30) in strains of Vibrio cholerae of O1/non-O1 serogroups of various origin, that is a component of chitinolytic complex taking into account object of isolation and epidemiologic significance of strains. MATERIALS AND METHODS: Cultures of V. cholerae O1/non-O1 serogroup strains were obtained from the museum of live culture of Rostov RIPC. Enzymatic activity analysis was carried out in Hitachi F-2500 fluorescent spectrophotometer using FL Solutions licensed software. NCBI databases were used during enzyme characteristics. RESULTS: N-acetyl-ß-D-glucosaminidase in Vcholerae O1/non-O1 serogroup strains was detected, purified by column chromatography, studied and characterized by a number of physical-chemical and biological properties. Comparative computer analysis of amino acid sequence of N-acetyl-ß-D-glucosaminidases of V. cholerae (VC2217 gene), Serratia marcescens etc. has allowed. to attribute the enzyme from V. cholerae to glycosyl-hydrolases (chitobiases) of family 20 and classify it according to enzyme nomenclature as EC 3.2.1.30. CONCLUSION: N-acetyl-ß-D-glucosaminidase in V. cholerae of O1/non-O1 serogroups of various origin and epidemiologic significance, participating in chitin utilization was studied and characterized for the first time, and its possible role in biology of cholera causative agent was shown.
Subject(s)
Acetylglucosaminidase/genetics , Cholera/enzymology , Vibrio cholerae O1/genetics , Vibrio cholerae non-O1/isolation & purification , Acetylglucosaminidase/classification , Amino Acid Sequence , Cholera/epidemiology , Cholera/microbiology , Humans , Serratia marcescens/enzymology , Vibrio cholerae O1/isolation & purification , Vibrio cholerae O1/pathogenicity , Vibrio cholerae non-O1/genetics , Vibrio cholerae non-O1/pathogenicityABSTRACT
Reviewed the paper are the composition and functions of Vibrio cholerae chitinolytic complex which play an important role in the maintaining and creating new forms of vibrios in the environ- ment, it is better adapted to survive in environmental.
Subject(s)
Adaptation, Physiological/physiology , Bacterial Proteins/metabolism , Chitinases/metabolism , Vibrio cholerae/enzymology , Animals , Bacterial Proteins/genetics , Chitinases/genetics , Humans , Vibrio cholerae/geneticsABSTRACT
AIM: Study system of activation of plasminogen in Vibrio cholerae. MATERIALS AND METHODS: 75 strains of V. cholerae of various origins were used in the study. Plasminogen was isolated from human plasma by using affinity chromatography on L-lysine sepharose, alpha-enolase activity was determined by a direct method assuming transformation of 2-phosphoglycerate into phopshoenolpyruvate. Vibrios were destroyed by ultrasound disintegrator to isolate membrane Omp protein, intact cells were discarded by centrifugation and cell lysate was centrifugated for 1 hour at 105000 g. The precipitate was solubilized in buffer with 1% triton X-100 and passed through a column with DE-52 cellulose. RESULTS: Vibrio cholerae O1 and O139 strains isolated from clinical specimens and water samples from open water bodies had the ability to bind by using alpha-enolase and transform human plasminogen into plasmin under the effect of outer membrane protein OmpT A protein with molecular weight around 40 kDa had proteolytic activity with a wide specter of substrate specificity, degraded fibrin, gelatin, collagen, protamine and activated plasminogen. Computer analysis showed that OmpT protein of cholera vibrion had a low degree of relation with Enterobacteriaceae omptins. CONCLUSION: The study carried out showed that vibrios have a system of activation of plasminogen that includes at least alpha-enolase and OmpT membrane protein. OmpT protein is assumed to belong to a new class of porins of Vibrionaceae family and its enzymatic activity may play a significant role in pathogenesis of infection.
Subject(s)
Bacterial Proteins/chemistry , Phosphopyruvate Hydratase/chemistry , Plasminogen/chemistry , Porins/chemistry , Proteolysis , Vibrio cholerae/enzymology , Bacterial Proteins/metabolism , Enzyme Activation , Humans , Phosphopyruvate Hydratase/metabolism , Plasminogen/metabolism , Porins/metabolismABSTRACT
The article discusses the technique of defining the strains of comma bacillus according their capability to convert human plasminogen into plasmin in vitro. This method can be implemented in applied and fundamental research concerning the study of subtle mechanisms of pathogenesis and colonization of intestines under cholera.
Subject(s)
Fibrinolysin/metabolism , Plasminogen/metabolism , Vibrio cholerae/physiology , Arginine/metabolism , Enzyme Activation , Humans , Hydrolysis , Spectrophotometry , Vibrio cholerae/isolation & purificationABSTRACT
Antiplague Research Institute, Rostov-on-Don, Russia Retrospective multi-locus VNTR-analysis was made for 166 Vibrio cholerae strains isolated, 1967-2001, in Rostov Region from clinical samples (82 strains) and from water samples (84 strains). On the basis of cluster analysis of heterogeneous identification strain genotypes, 45 variations of individual strains were shared between 11 separate clusters, among which the F cluster vibrios were predominant. Having emerged, 1970, in the region, they were widely spread during the 1973-1975 cholera pandemic and were registered, among the isolated strains, till 1992 indicating the possibility of long persistence of V. cholerae 01 in the natural aquatic environment. Presumably, the ecosystem specificity contributed to the long-term vibrio persistence.
Subject(s)
Cholera/virology , Disease Outbreaks , Vibrio cholerae O1/genetics , Water Microbiology , Alleles , Cholera/epidemiology , Cluster Analysis , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Fresh Water/microbiology , Genotype , Humans , Molecular Epidemiology , Retrospective Studies , Russia/epidemiology , Sequence Analysis, DNA , Tandem Repeat Sequences , Vibrio cholerae O1/isolation & purificationABSTRACT
The comparative study of variable tandem repeats (VNTR analysis) in genomes of V. cholerae 0139 isolated from humans and from water samples taken from surface reservoirs was carried out. The results of the study of the allele state of 5 loci of tandem repeats in 50 strains of vibrios, carried out in the double-primer polymerase chain reaction (PCR), as well as the earlier comparison of the same isolates in the single-primer PCR, showed essential differences and the absence of clonality in the cultures of the clinical and aqueous origin. The suggestion was made that vibrios with individual VNTR genotypes and having no genes ctx and tcpA, isolated from water samples, were epidemic unimportant representatives of the autochthonous microflora of water reservoirs.
Subject(s)
Genes, Bacterial , Tandem Repeat Sequences , Vibrio cholerae O139/genetics , Water Microbiology , Alleles , Cholera/epidemiology , Cholera Toxin/genetics , Fimbriae Proteins/genetics , Fresh Water/microbiology , Genotype , Humans , Polymerase Chain Reaction , Russia/epidemiology , Vibrio cholerae O139/isolation & purificationABSTRACT
Computer analysis revealed seven potential variable-number tandem-repeat (VNTR) loci in the Vibrio cholerae genome. Specific primers were designed to amplify locus VcA located on chromosome 2 and containing a TGCTGT repeat. The locus was found in all tested strains from a V. cholerae strain collection, the repeat number varying 3 to 23. In total, 14 VcA alleles were observed. The VcA locus was proposed as a marker for the molecular typing of V. cholerae strains.
Subject(s)
Minisatellite Repeats , Vibrio cholerae/genetics , Algorithms , Alleles , Bacterial Typing Techniques/methods , Base Sequence , DNA Primers , Databases, Nucleic Acid , Molecular Sequence Data , Polymerase Chain Reaction/methods , Vibrio cholerae/classificationABSTRACT
The comparative study of the genomes of V. cholerae O139 isolated from humans and from water of surface reservoirs was carried out with the use of single- and double-primer polymerase chain reaction (PCR). The profiles of polymorphic DNA fragments obtained in this study made it possible to find out differences between groups of strains, as well as the individual features of some of them. The comparison of strains isolated from humans and from water in single-primer PCR revealed that they, in spite of the general similarity of their genomes, essentially differed, which was probably due to changes in the genome of this infective agent. Strains of aqueous origin lacked genes ctx and tcpA, which made them epidemiologically unimportant.
