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 in Swiss-type cheese. In this study, 68 strains of P. freudenreichii were screened for the beta-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 beta-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 beta-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 beta-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 beta-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.
Deutsch
Stéphanie-Marie
SM
INRA, UMR1253 Science et Technologie du Lait et de l'OEuf, Rennes, France. stephanie-marie.deutsch@rennes.inra.fr
Le Bivic
Pierre
P
Hervé
Christophe
C
Madec
Marie-Noëlle
MN
LaPointe
Gisèle
G
Jan
Gwenaël
G
Le Loir
Yves
Y
Falentin
Hélène
H
D016667:Bacterial Capsules / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D015870:Gene Expression; D005798:Genes, Bacterial; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011424:Propionibacterium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D047071:beta-Glucans / Q000032:analysis* / Q000378:metabolism
D047071:beta-Glucans; D016695:Glycosyltransferases