Specific metabolic activity of ripening bacteria quantified by real-time reverse transcription PCR throughout Emmental cheese manufacture

Falentin, Hélène; Postollec, Florence; Parayre, Sandrine; Henaff, Nadine; Bivic, Pierre Le; Richoux, Romain; Thierry, Anne; Sohier, Danièle

doi:10.1016/j.ijfoodmicro.2010.06.003

Cited by 67 publications

(50 citation statements)

References 34 publications

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“…To follow gene expression of P. freudenreichii and Lb. paracasei during cheese-making, Falentin et al (2010) measured the amount of cDNA copies of the target sequence after reverse transcription, and divided this value by the corresponding number of cells, which was measured by qPCR analysis of DNA extracted from the cheese samples. From these analyses, it could be concluded that the metabolic activity of Lb.…”

Section: Real-time Pcr Methodsmentioning

confidence: 99%

“…RNA extraction methods involving prior separation of the cells from cheeses and other dairy products have been used in several studies (Randazzo et al, 2002;Bleve et al, 2003;Sanchez et al, 2006;Bogovic Matijasic et al, 2007;Smeianov et al, 2007;Makhzami et al, 2008;Rantsiou et al, 2008a;Rantsiou et al, 2008b;Ulvé et al, 2008;Duquenne et al, 2010;Falentin et al, 2010;Cretenet et al, 2011;La Gioia et al, 2011;Masoud et al, 2011;Taïbi et al, 2011). The recovery of microbial cells is done following similar protocols than for DNA extraction methods (see above).…”

Section: Rna Extractionmentioning

confidence: 99%

“…Propionibacterium (P.) freundereichii, a species involved in Emmental cheese ripening, has a thick cell wall surrounded with capsular exopolysaccharides. For an efficient lysis of P. freundereichii cells recovered from cheeses, Falentin et al (2010) used a combination of lysozyme treatment, bead-beating and phenol-chloroform extraction. Sanchez et al (2006) recovered lactic acid bacteria cells from milk cultures after dispersion of caseins with EDTA, and extracted RNA using guanidinium thiocyanate-phenol-chloroform (commercial TRIzol reagent), a reagent that inactivates cellular processes and allows separation of RNA from DNA and proteins (Chomczynski and Sacchi, 1987).…”

Section: Rna Extractionmentioning

confidence: 99%

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Application of PCR-Based Methods to Dairy Products and to Non-Dairy Probiotic Products

Monnet¹,

Matijašić²

2012

Polymerase Chain Reaction

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Section: Real-time Pcr Methodsmentioning

confidence: 99%

Section: Rna Extractionmentioning

confidence: 99%

Section: Rna Extractionmentioning

confidence: 99%

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Application of PCR-Based Methods to Dairy Products and to Non-Dairy Probiotic Products

Monnet¹,

Matijašić²

2012

Polymerase Chain Reaction

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“…Ptuf is the upstream region of the tuf gene (236 bp) of P. freudenreichii CB1 (accession number FN 806773) that encodes the translation elongation factor. This gene was shown to be strongly expressed during the growth of the strain in YEL (18). To perform the construction, Ptuf and gtfF were amplified by PCR from P. freudenreichii CB1 genomic DNA using the primers ptuf-bamH1-F/ptuf-xba1-R and gtfF-XbaI-F/gtfF-HindIII-R (see Table S1 in the supplemental material).…”

Section: Construction Of the Genetically Modified Strains (I) Insertmentioning

confidence: 99%

Contribution of Surface β-Glucan Polysaccharide to Physicochemical and Immunomodulatory Properties of Propionibacterium freudenreichii

Deutsch

Parayre

Bouchoux

et al. 2012

Appl Environ Microbiol

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e Propionibacterium freudenreichii is a bacterial species found in Swiss-type cheeses and is also considered for its health properties. The main claimed effect is the bifidogenic property. Some strains were shown recently to display other interesting probiotic potentialities such as anti-inflammatory properties. About 30% of strains were shown to produce a surface exopolysaccharide (EPS) composed of (1¡3,1¡2)-␤-D-glucan due to a single gene named gtfF. We hypothesized that functional properties of P. freudenreichii strains, including their anti-inflammatory properties, could be linked to the presence of ␤-glucan. To evaluate this hypothesis, gtfF genes of three ␤-glucan-producing strains were disrupted. These knockout (KO) mutants were complemented with a plasmid harboring gtfF (KO-C mutants). The absence of ␤-glucan in KO mutants was verified by immunological detection and transmission electron microscopy. We observed by atomic force microscopy that the absence of ␤-glucan in the KO mutant dramatically changed the cell's topography. The capacity to adhere to polystyrene surface was increased for the KO mutants compared to wild-type (WT) strains. Antiinflammatory properties of WT strains and mutants were analyzed by stimulation of human peripheral blood mononuclear cells (PBMCs). A significant increase of the anti-inflammatory interleukin-10 cytokine production by PBMCs was measured in the KO mutants compared to WT strains. For one strain, the role of ␤-glucan in mice gut persistence was assessed, and no significant difference was observed between the WT strain and its KO mutant. Thus, ␤-glucan appears to partly hide the anti-inflammatory properties of P. freudenreichii; which is an important result for the selection of probiotic strains.

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“…It grows in cheese during ripening at warm temperatures (20 to 24°C) but remains metabolically active during the storage of cheese at low temperatures (10). We previously investigated the adaptation strategies of P. freudenreichii type strain CIRM-BIA1…”

mentioning

confidence: 99%

Accumulation of Intracellular Glycogen and Trehalose by Propionibacterium freudenreichii under Conditions Mimicking Cheese Ripening in the Cold

Dalmasso

Aubert

Even

et al. 2012

Appl Environ Microbiol

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f Seven Propionibacterium freudenreichii strains exhibited similar responses when placed at 4°C. They slowed down cell machinery, displayed cold stress responses, and rerouted their carbon metabolism toward trehalose and glycogen synthesis, both accumulated in cells. These results highlight the molecular basis of long-term survival of P. freudenreichii in the cold. Propionibacterium freudenreichii is a bacterium of food and probiotic interest, widely used as a ripening culture in the manufacture of Swiss cheese varieties (4, 16). It grows in cheese during ripening at warm temperatures (20 to 24°C) but remains metabolically active during the storage of cheese at low temperatures (10). We previously investigated the adaptation strategies of P. freudenreichii type strain CIRM-BIA1T by -omic approaches under conditions mimicking cheese ripening in the cold (6). Our previous results suggest in particular that CIRM-BIA1T reroutes its metabolism toward glycogen synthesis. In the present study, we confirmed the actual accumulation of glycogen in cells and investigated the response in the cold of six other P. freudenreichii strains.Choice of strains and culture conditions. The transcriptomic response of six P. freudenreichii subsp. shermanii strains (CIRM-BIA9, CIRM-BIA118, CIRM-BIA122, and CIRM-BIA123 from CIRM-BIA [Centre International de Ressources MicrobiennesBactéries d'Intérêt Alimentaire, INRA, Rennes, France] and CIRM-BIA472 and CIRM-BIA482 from Valio Ltd., Helsinki, Finland) was studied during their transfer from 30°C to 4°C under conditions mimicking cheese ripening, previously applied to strain CIRM-BIA1 T (6). All experiments were made in triplicate independent cultures. The six strains were chosen with different sequence types (7) and phenotypes. For example, they produce methylbutanoate and ethyl propionate, two cheese aroma compounds, at concentrations varying by factors of 6 and 12, respectively, depending on the strain (data not shown).Growth and metabolite production in the cold. All the strains stopped their growth when placed at 4°C, whereas in the control cultures maintained at 30°C, cells went on growing for about 20 h (Fig. 1A). They went on producing propionate and acetate, the two main products of lactate fermentation, but at a markedly lower production rate in the cold (Fig. 1C and D) (3.4 Ϯ 0.6 [mean Ϯ standard deviation] mM per day at 4°C versus 76 Ϯ 15 mM per day at 30°C, i.e., a 23-Ϯ 6-fold decrease for propionate). The rate of methylbutanoate production also decreased but at a markedly lower extent (from 69 Ϯ 55 M per day at 30°C to 12 Ϯ 12 M per day at 4°C, i.e., a mean fold decrease of 7 Ϯ 4) (Fig. 1B).Transcriptomic approach applied to all strains. Gene expression after an 80-h period at 4°C (t ϭ 120 h) was compared to that at 20 h during growth at 30°C for the 6 strains, using the methodology and microarrays previously described for strain CIRM- BIA1T (6) (NCBI GEO, http://www.ncbi.nlm.nih.gov/geo/, platform accession number GPL13959). The transcriptomic data for

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Specific metabolic activity of ripening bacteria quantified by real-time reverse transcription PCR throughout Emmental cheese manufacture

Cited by 67 publications

References 34 publications

Application of PCR-Based Methods to Dairy Products and to Non-Dairy Probiotic Products

Application of PCR-Based Methods to Dairy Products and to Non-Dairy Probiotic Products

Contribution of Surface β-Glucan Polysaccharide to Physicochemical and Immunomodulatory Properties of Propionibacterium freudenreichii

Accumulation of Intracellular Glycogen and Trehalose by Propionibacterium freudenreichii under Conditions Mimicking Cheese Ripening in the Cold

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