Correlation of the Capsular Phenotype in Propionibacterium freudenreichii with the Level of Expression of gtf, a Unique Polysaccharide Synthase-Encoding Gene

Many food-grade bacteria produce exopolysaccharides (EPS) that affect the texture of fermented food products and that may be involved in probiotic properties. Propionibacterium freudenreichii is a Gram-positive food-grade bacterium with reported probiotic capabilities that is widely used as starter...

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Veröffentlicht in:Applied and Environmental Microbiology 2010-05, Vol.76 (9), p.2740-2746
Hauptverfasser: Deutsch, Stéphanie-Marie, Bivic, Pierre Le, Hervé, Christophe, Madec, Marie-Noëlle, LaPointe, Gisèle, Jan, Gwenaël, Loir, Yves Le, Falentin, Hélène
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container_end_page 2746
container_issue 9
container_start_page 2740
container_title Applied and Environmental Microbiology
container_volume 76
creator Deutsch, Stéphanie-Marie
Bivic, Pierre Le
Hervé, Christophe
Madec, Marie-Noëlle
LaPointe, Gisèle
Jan, Gwenaël
Loir, Yves Le
Falentin, Hélène
description Many food-grade bacteria produce exopolysaccharides (EPS) that affect the texture of fermented food products and that may be involved in probiotic properties. Propionibacterium freudenreichii is a Gram-positive food-grade bacterium with reported probiotic capabilities that is widely used as starter in Swiss-type cheese. In this study, 68 strains of P. freudenreichii were screened for the β-glucan capsular phenotype by immunoagglutination with a specific antibody and for the presence of the gtf gene coding for polysaccharide synthase. All strains were positive for PCR amplification with gtf gene-specific primers, but the presence of β-glucan capsular EPS was detected for only 35% of the strains studied. Disruption of gtf in P. freudenreichii revealed that gtf is a unique gene involved in β-glucan capsular EPS production in P. freudenreichii. The gtf gene was transferred into and expressed in Lactococcus lactis, in which it conferred an agglutination-positive phenotype. Expression of the gtf gene was measured by performing quantitative reverse transcription-PCR assays with RNA from four capsular and three noncapsular strains. A positive correlation was found between the β-glucan capsular phenotype and gtf gene expression. Sequencing of the region upstream of the gtf open reading frame revealed the presence of an insertion element (IS element) in this upstream region in the four strains with the β-glucan capsular phenotype. The role of the IS element in the expression of neighboring genes and its impact on interstrain variability of the P. freudenreichii capsule phenotype remain to be elucidated.
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Propionibacterium freudenreichii is a Gram-positive food-grade bacterium with reported probiotic capabilities that is widely used as starter in Swiss-type cheese. In this study, 68 strains of P. freudenreichii were screened for the β-glucan capsular phenotype by immunoagglutination with a specific antibody and for the presence of the gtf gene coding for polysaccharide synthase. All strains were positive for PCR amplification with gtf gene-specific primers, but the presence of β-glucan capsular EPS was detected for only 35% of the strains studied. Disruption of gtf in P. freudenreichii revealed that gtf is a unique gene involved in β-glucan capsular EPS production in P. freudenreichii. The gtf gene was transferred into and expressed in Lactococcus lactis, in which it conferred an agglutination-positive phenotype. Expression of the gtf gene was measured by performing quantitative reverse transcription-PCR assays with RNA from four capsular and three noncapsular strains. 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subjects Antibodies
Bacterial Capsules - chemistry
Bacterial Capsules - metabolism
Base Sequence
beta-Glucans - analysis
beta-Glucans - metabolism
Carbohydrates
Ecology, environment
Gene Expression
Genes, Bacterial
Genetics and Molecular Biology
Genotype & phenotype
Glycosyltransferases - genetics
Glycosyltransferases - metabolism
Gram-positive bacteria
Immunoglobulins
Lactococcus lactis
Life Sciences
Microbiology
Molecular Sequence Data
Phenotype
Polymerase Chain Reaction
Probiotics
Propionibacterium - enzymology
Propionibacterium - genetics
Propionibacterium - metabolism
title Correlation of the Capsular Phenotype in Propionibacterium freudenreichii with the Level of Expression of gtf, a Unique Polysaccharide Synthase-Encoding Gene
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