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Exopolysaccharide production by lactic acid bacteria is beneficial in the dairy and oat-based food industries and is used to improve the texture of the fermented products. However, β-D-glucan–producing bacteria are considered spoilage microorganisms in alcoholic beverages
because their secreted exopolysaccharides alter the viscosity of cider, wine, and beer, rendering them unpalatable. The plasmidic glycosyltransferase (gtf) gene of the Pediococcus parvulus 2.6 strain isolated from ropy cider has been cloned and sequenced, and its GTF product
was functionally expressed in Streptococcus pneumoniae. The GTF protein, which has glycosyltransferase activity, belongs to the COG1215 membrane-bound glycosyltransferase family, and agglutination tests revealed that the enzyme enables S. pneumoniae to synthesize β-D-glucan.
PCR amplification and Southern blot hybridization showed that the gtf gene is also present at different genomic locations in the β-D-glucan producers Lactobacillus diolivorans G77 and Oenococcus oeni I4 strains, also isolated from ropy cider. A PCR assay has been
developed for the detection of exopolysaccharide-producing bacteria. Forward and reverse primers, included respectively in the coding sequences of the putative glycosyltransferase domain and the fifth trans-membrane segment of the GTF, were designed. Analysis of 76 ropy and nonropy
lactic acid bacteria validated the method for specific detection of β-D-glucan homopolysaccharide producer Pediococcus, Lactobacillus, and Oenococcus strains. The limit of the assay in cider was 3 × 102 CFU/ml. This molecular method can be useful for the
detection of ropy bacteria in cider before spoilage occurs, as well as for isolation of new exopolysaccharide-producing strains of industrial interest.
Document Type: Research Article
Departamento de Estructura y Función de proteínas, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain 2:
Departamento de Quimica Aplicada, Facultad de Ciencias Quimicas, Box 1072, 20080 San Sebastian, Spain 3:
Departamento de Biologia Molecular (Unidad Asociada al Centro de Investigaciones Biologicas, C.S.I.C.), Facultad de Medicina, Universidad de Cantabria, 39011 Santander, Spain
Publication date: January 1, 2006
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