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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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. 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.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>EISSN: 1098-6596</identifier><identifier>DOI: 10.1128/AEM.02591-09</identifier><identifier>PMID: 20228100</identifier><identifier>CODEN: AEMIDF</identifier><language>eng</language><publisher>United States: American Society for Microbiology</publisher><subject>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</subject><ispartof>Applied and Environmental Microbiology, 2010-05, Vol.76 (9), p.2740-2746</ispartof><rights>Copyright American Society for Microbiology May 2010</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><rights>Copyright © 2010, American Society for Microbiology 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c569t-d93b4553af8d8d953cc44f9d5422fcd6260d9a2ef61eb77dafa293fadeeb8b313</citedby><cites>FETCH-LOGICAL-c569t-d93b4553af8d8d953cc44f9d5422fcd6260d9a2ef61eb77dafa293fadeeb8b313</cites><orcidid>0000-0001-8694-3381 ; 0000-0002-0355-1065 ; 0000-0002-5570-5193 ; 0000-0001-6254-5303</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2863425/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2863425/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,3175,3176,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20228100$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.inrae.fr/hal-02655923$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Deutsch, Stéphanie-Marie</creatorcontrib><creatorcontrib>Bivic, Pierre Le</creatorcontrib><creatorcontrib>Hervé, Christophe</creatorcontrib><creatorcontrib>Madec, Marie-Noëlle</creatorcontrib><creatorcontrib>LaPointe, Gisèle</creatorcontrib><creatorcontrib>Jan, Gwenaël</creatorcontrib><creatorcontrib>Loir, Yves Le</creatorcontrib><creatorcontrib>Falentin, Hélène</creatorcontrib><title>Correlation of the Capsular Phenotype in Propionibacterium freudenreichii with the Level of Expression of gtf, a Unique Polysaccharide Synthase-Encoding Gene</title><title>Applied and Environmental Microbiology</title><addtitle>Appl Environ Microbiol</addtitle><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.</description><subject>Antibodies</subject><subject>Bacterial Capsules - chemistry</subject><subject>Bacterial Capsules - metabolism</subject><subject>Base Sequence</subject><subject>beta-Glucans - analysis</subject><subject>beta-Glucans - metabolism</subject><subject>Carbohydrates</subject><subject>Ecology, environment</subject><subject>Gene Expression</subject><subject>Genes, Bacterial</subject><subject>Genetics and Molecular Biology</subject><subject>Genotype & phenotype</subject><subject>Glycosyltransferases - genetics</subject><subject>Glycosyltransferases - metabolism</subject><subject>Gram-positive bacteria</subject><subject>Immunoglobulins</subject><subject>Lactococcus lactis</subject><subject>Life Sciences</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Phenotype</subject><subject>Polymerase Chain Reaction</subject><subject>Probiotics</subject><subject>Propionibacterium - enzymology</subject><subject>Propionibacterium - genetics</subject><subject>Propionibacterium - metabolism</subject><issn>0099-2240</issn><issn>1098-5336</issn><issn>1098-6596</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkl1v0zAUhiMEYmVwxzUYbhDSMvyRL98gVVXZkIqoNHptOfZJ4ymxMzvp6I_hv5K0ZYLdcGXJfs5z7OM3il4TfEkILT7Nl98uMU05iTF_Es0I5kWcMpY9jWYYcx5TmuCz6EUItxjjBGfF8-iMYkoLgvEs-rVw3kMje-MschXqa0AL2YWhkR6ta7Cu33eAjEVr77oRMqVUPXgztKjyMGiwHoyqjUH3pq8P9SvYQTPJlj87DyGc1Nu-ukASbay5GwCtXbMPUqlaeqMB3extX8sA8dIqp43doiuw8DJ6VskmwKvTeh5tvix_LK7j1ferr4v5KlZpxvtYc1YmacpkVehC85QplSQV12lCaaV0RjOsuaRQZQTKPNeykpSzSmqAsigZYefR56O3G8oWtALbe9mIzptW-r1w0oh_T6ypxdbtBC0yltB0FHw8CupHZdfzlZj2MM3SlFO2m5p9ODXzbpxE6EVrgoKmkRbcEESeJFnORu3_ScYYwcWBfP-IvHWDt-PMBMUpzxNOJujiCCnvQvBQPdyUYDFlSYxZEocsCcxH_M3fQ3mA_4RnBN6dHm229b3xIGRohYRW5JnggubJxLw9MpV0Qm69CWJzQzFhmBSMFONf_Qbkctus</recordid><startdate>20100501</startdate><enddate>20100501</enddate><creator>Deutsch, Stéphanie-Marie</creator><creator>Bivic, Pierre Le</creator><creator>Hervé, Christophe</creator><creator>Madec, Marie-Noëlle</creator><creator>LaPointe, Gisèle</creator><creator>Jan, Gwenaël</creator><creator>Loir, Yves Le</creator><creator>Falentin, Hélène</creator><general>American Society for Microbiology</general><general>American Society for Microbiology (ASM)</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>SOI</scope><scope>7X8</scope><scope>1XC</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8694-3381</orcidid><orcidid>https://orcid.org/0000-0002-0355-1065</orcidid><orcidid>https://orcid.org/0000-0002-5570-5193</orcidid><orcidid>https://orcid.org/0000-0001-6254-5303</orcidid></search><sort><creationdate>20100501</creationdate><title>Correlation of the Capsular Phenotype in Propionibacterium freudenreichii with the Level of Expression of gtf, a Unique Polysaccharide Synthase-Encoding Gene</title><author>Deutsch, Stéphanie-Marie ; 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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. 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.</abstract><cop>United States</cop><pub>American Society for Microbiology</pub><pmid>20228100</pmid><doi>10.1128/AEM.02591-09</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0001-8694-3381</orcidid><orcidid>https://orcid.org/0000-0002-0355-1065</orcidid><orcidid>https://orcid.org/0000-0002-5570-5193</orcidid><orcidid>https://orcid.org/0000-0001-6254-5303</orcidid><oa>free_for_read</oa></addata></record> |
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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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