Identification and characterisation of probiotic lactic acid bacteria

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Authors

ŠPANOVÁ Alena RITTICH Bohuslav HORÁK Daniel DRÁB Vladimír DRBOHLAV Jan

Year of publication 2008
Type Article in Proceedings
Conference 3rd International Congress on Bioprocesses in Food Industries (ICBF 2008)
MU Faculty or unit

Faculty of Science

Citation
Field Genetics and molecular biology
Keywords magnetic particles; P(HEMA-co-EDMA); P(HEMA-coGMA); PGMA; immobilization; enzymes; DNA;
Description Polyphasic approach was applied for lactic acid bacteria isolated from suckling babies and healthy adults and lactic acid bacteria collected in the Culture Collection of Dairy Microorganisms (Laktoflora, Tábor, Czech Republic). Altogether 80 strains of the Bifidobacterium genus and 60 strains of the Lactobacillus genus were tested. Some functional properties of analysed strains (such as surviving at low pH, resistance to bile salts, growth in milk, stability during storage, or influence on the growth of selected starter cultures used in milk industry) were proved. Several molecular biology methods (genus-specific PCRs, species-specific PCRs, amplified ribosomal DNA restriction analysis (ARDRA), RAPDs, and rep-PCRs) were used for the identification and characterisation of bacterial strains. Attention was concentrated on strains identified in human probiotic species. Magnetic nonporous hydrophilic microspheres poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) - (P(HEMA-co-EDMA) and poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) - P(HEMA-co-GMA) containing carboxyl groups were used for whole DNA isolation from different types of complex food samples (liquid milk products, yoghurts, cheeses) containing PCR inhibitors. Genomic DNA was successfully purified from cell lysates using tested microspheres in the presence of PEG and sodium chloride. The purified DNA was suitable for the amplification and identification of studied bacterial strains in PCR-based methods.Genes coding for bacteriocins were identified using DNA probe purified with the help of magnetic microspheres in more Lactobacillus and Lactococcus lactis strains with success.
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