Food Microbiology

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

Marion Dalmasso, Julie Aubert, Sergine Even, Hélène Falentin, Marie-Bernadette Maillard, Sandrine Parayre, Valentin Loux, Jarna Tanskanen, Anne Thierry
DOI: 10.1128/AEM.00561-12

Article Figures & Data

Figures

  • Fig 1
    Fig 1

    Time course of metabolic activity of seven P. freudenreichii strains over a 40-h incubation at 30°C followed by a further 80 h at 4°C. Growth (OD650nm, optical density at 650 nm) (A), concentrations of methylbutanoate (sum of 2-methylbutanoate and 3-methylbutanoate) (B), propionate (C), and acetate (D). Error bars show the standard deviations of the results of triplicate independent experiments. The inset in panel A shows the growth curves at 4°C and 30°C. Values are means for the 7 strains: CIRM-BIA1T (×), CIRM-BIA9 (△), CIRM-BIA118 (□), CIRM-BIA122 (▲), CIRM-BIA123 (○), CIRM-BIA472 (●), CIRM-BIA482 (■).

  • Fig 2
    Fig 2

    Number of differentially expressed genes (|fold change| > 1) after 80 h at 4°C in comparison with gene expression at the reference time of 20 h for seven P. freudenreichii strains (CIRM-BIA118, -122, -123, -1, -472, -482, and -9). Downregulated (white bars) or upregulated (black bars) genes with known functions are presented according to their metabolic category: E, energy metabolism; P, protein synthesis; T, transport of peptides and inorganic ions; AA, transport and metabolism of amino acids; CH, transport and metabolism of carbohydrates; A, adaptation to atypical conditions.

  • Fig 3
    Fig 3

    Main routes of pyruvate formation and conversion in P. freudenreichii during storage at 4°C and relevant for this study. Genes upregulated at 4°C are shown in black, and downregulated genes in gray. Thick black arrows emphasize the metabolic pathways that are favored at 4°C, and thin gray arrows the pathways that are downregulated at 4°C. ace, pyruvate dehydrogenase, E1 component; ald, alanine dehydrogenase; ldh, l-lactate dehydrogenase; ppdk, pyruvate phosphate dikinase; sdaA, l-serine dehydratase; sdh, succinate dehydrogenase; CoA, coenzyme A. Values of fold changes are shown in Table 1, Table 4, and Fig. 5.

  • Fig 4
    Fig 4

    Changes in expression of genes involved in trehalose and glycogen synthesis in P. freudenreichii (pathways adapted from Chandra et al. [3]). Each box shows the fold change, expressed as log2, for each gene in all seven strains (CIRM-BIA118, -122, -123, -1, -472, -482, and -9) after 80 h at 4°C in comparison with gene expression at the reference time (20 h). Values of |fold change (log2)| >1 and <−1 are shown in dark and light gray, respectively.

  • Fig 5
    Fig 5

    Accumulation of intracellular glycogen and trehalose in P. freudenreichii CIRM-BIA1T during growth at 30°C (up to 40 h of incubation) and further incubation for 250 h at 4°C. Values are means of the results of triplicate independent experiments; error bars show standard deviations. eq, equivalent.

Tables

  • Table 1

    Differentially expressed genes involved in general cell machinery slowdown

    NameLocus tagaDescriptionCategorybP valueFold change (log2) for CIRM-BIA strainc:
    TimeStrainTime × strain11812212314724829
    cstAPFREUD_16500Carbon starvation proteinA<0.01<0.01<0.01−1.0−1.2−3.4−1.5−2.0−3.7−2.9
    ftsXPFREUD_09600Cell division proteinCD<0.01<0.01<0.01−2.1−1.7−2.4−4.7−1.7−3.8−4.3
    icdPFREUD_06870Putative isocitrate/isopropylmalate dehydrogenaseCH<0.01<0.010.05−1.0−3.3−2.0−3.0−3.4−3.3−3.0
    pccBPFREUD_07170Propionyl-coenzyme A carboxylase beta chainCH<0.010.01<0.01−1.2−1.6−0.4−1.8−1.8−1.5−1.2
    aceEPFREUD_09470Pyruvate dehydrogenase E1 componentCH<0.01<0.010.14−1.5−1.9−1.5−1.9−2.8−2.1−1.9
    lpdPFREUD_10890Dihydrolipoyl dehydrogenaseCH<0.010.280.44−0.5−0.9−0.7−1.5−2.2−0.8−1.1
    acnPFREUD_12590AconitaseCH<0.01<0.010.47−0.7−0.8−1.0−1.0−1.5−0.8−1.0
    cydAPFREUD_01720Cytochrome d ubiquinol oxidase, subunit IE<0.01<0.010.02−0.8−1.4−1.6−1.6−1.5−2.0−3.4
    cydBPFREUD_01730Cytochrome d ubiquinol oxidase, subunit IIE<0.01<0.010.03−1.0−1.3−1.3−1.1−0.7−0.9−2.2
    nuoAPFREUD_05160NADH-quinone oxidoreductase chain AE<0.01<0.010.02−1.4−1.2−0.5−2.0−1.3−2.2−2.0
    nuoBPFREUD_05170NADH-quinone oxidoreductase chain BE<0.01<0.01<0.01−1.9−1.8−1.0−2.3−1.4−3.3−2.3
    nuoCPFREUD_05180NADH-quinone oxidoreductase chain CE<0.01<0.01<0.01−1.9−1.7−1.3−3.8−2.3−3.7−3.4
    nuoDPFREUD_05190NADH-quinone oxidoreductase chain DE<0.01<0.01<0.01−2.0−1.9−1.6−3.8−2.7−4.9−5.2
    nuoEPFREUD_05200NADH-quinone oxidoreductase chain EE<0.01<0.01<0.01−2.0−1.9−1.9−2.9−2.5−4.0−3.3
    nuoFPFREUD_05210NADH-quinone oxidoreductase chain FE<0.01<0.010.08−2.2−2.0−1.9−2.7−2.4−3.4−2.9
    nuoGPFREUD_05220NADH-quinone oxidoreductase chain GE<0.01<0.010.08−2.0−1.5−1.9−1.9−2.2−2.8−3.1
    nuoHPFREUD_05230NADH-quinone oxidoreductase chain HE<0.01<0.010.01−1.8−1.0−1.6−0.9−1.6−2.4−1.7
    nuoIPFREUD_05240NADH-quinone oxidoreductase chain IE<0.01<0.010.09−2.5−1.0−1.6−1.4−2.3−3.0−2.9
    nuoJPFREUD_05250NADH-quinone oxidoreductase chain JE<0.01<0.010.02−2.1−1.1−1.7−1.5−2.0−3.3−2.1
    nuoKPFREUD_05260NADH dehydrogenase I chain KE<0.01<0.010.01−2.3−1.2−1.5−2.2−2.1−3.4−2.6
    nuoLPFREUD_05270NADH dehydrogenaseE<0.01<0.010.01−2.5−1.5−2.1−1.9−2.5−3.7−2.9
    nuoMPFREUD_05280NADH dehydrogenase I chain ME<0.01<0.010.01−2.3−1.1−1.7−1.3−2.4−3.2−2.9
    nuoNPFREUD_05290NADH dehydrogenase I chain NE<0.01<0.010.05−2.8−1.6−2.1−1.2−2.3−2.8−2.3
    sdhC1PFREUD_09240Succinate dehydrogenase, subunit CE<0.01<0.01<0.01−1.3−0.81.2−2.1−2.2−2.1−2.2
    sdhAPFREUD_09250Succinate dehydrogenase, subunit AE<0.01<0.01<0.01−2.1−1.20.5−2.7−2.9−3.2−3.4
    sdhBPFREUD_09260Succinate dehydrogenase, subunit BE<0.01<0.010.09−1.2−0.90.3−0.9−1.0−0.7−1.1
    atpBPFREUD_10430ATP synthase A chainE<0.01<0.010.03−2.0−1.6−1.5−2.8−2.6−3.3−2.5
    atpEPFREUD_10440ATP synthase C chainE<0.01<0.010.22−2.8−2.7−1.9−3.1−3.1−2.8−3.2
    atpFPFREUD_10450ATP synthase B chainE<0.01<0.010.30−2.9−3.0−2.1−2.3−2.7−2.7−2.6
    atpHPFREUD_10460ATP synthase delta chainE<0.010.250.06−2.8−2.8−2.4−3.4−3.5−3.0−3.6
    atpAPFREUD_10470ATP synthase subunit alphaE<0.010.340.21−3.0−2.6−2.5−3.6−3.6−3.7−3.5
    atpGPFREUD_10480ATP synthase gamma chainE<0.01<0.010.20−3.1−2.6−2.4−3.7−4.2−3.9−3.5
    atpDPFREUD_10490ATP synthase subunit betaE<0.010.050.38−2.9−2.3−2.2−3.0−3.2−3.1−3.2
    atpCPFREUD_10500ATP synthase epsilon chainE<0.010.120.23−3.8−2.5−2.4−3.0−3.6−3.2−3.8
    sdhB3PFREUD_14300Succinate dehydrogenaseE<0.01<0.010.02−2.8−2.6−0.6−2.4−2.4−2.7−4.2
    sdhA3PFREUD_14310Succinate dehydrogenase flavoprotein subunitE<0.01<0.01<0.01−2.5−2.5−0.3−3.0−2.2−2.6−4.3
    sdhC2PFREUD_14320Succinate dehydrogenase cytochrome B-558 subunitE<0.01<0.01<0.01−2.4−2.20.3−2.3−1.5−2.3−2.8

    • ↵a Locus tag for CIRM-BIA1T.

    • ↵b A, adaptation to atypical conditions; CD, cell division; CH, transport and metabolism of carbohydrates; E, energy metabolism.

    • ↵c Values of |fold change (log2)| >1 are in boldface.

  • Table 2

    Differentially expressed genes involved in cold stress response

    NameLocus tagaDescriptionCategorybP valueFold change (log2) for CIRM-BIA strainc:
    TimeStrainTime × strain11812212314724829
    pspCPFREUD_06710Possible stress response transcriptional regulator proteinA<0.01<0.01<0.013.82.02.93.52.64.34.0
    pspCPFREUD_06710Possible stress response transcriptional regulator proteinA<0.01<0.01<0.013.82.02.93.52.64.34.0
    cspAPFREUD_09800Cold shock-like proteinA<0.01<0.010.01−0.30.11.62.11.31.71.9
    cspBPFREUD_18210Cold shock proteinA<0.01<0.010.010.80.61.12.30.81.80.9
    PFREUD_04260DeaD/DeaH box helicaseDNA<0.01<0.010.210.30.70.71.20.91.00.8
    PFREUD_13460Superfamily II RNA helicase, DeaD/DeaH box helicaseDNA<0.01<0.010.041.31.21.52.00.91.91.8
    dnaK2PFREUD_04630Chaperone proteinPM<0.01<0.01<0.01−0.5−1.8−2.6−2.5−2.5−0.50.5
    grpE2PFREUD_04640Co-chaperone proteinPM<0.010.04<0.010.9−0.9−1.5−2.4−1.50.00.9
    dnaJ2PFREUD_04650Chaperone protein DnaJ2PM<0.01<0.01<0.01−0.3−0.9−1.4−1.1−0.70.40.6
    groS1PFREUD_0646010-kDa chaperonin 1PM<0.010.020.01−2.4−4.1−3.9−5.5−5.4−5.9−5.2
    groL1PFREUD_0647060-kDa chaperonin 1PM<0.010.240.22−3.0−3.9−3.8−4.0−4.6−4.9−4.7
    groS2PFREUD_0781010-kDa chaperonin 2PM<0.01<0.010.15−0.8−0.60.3−0.7−1.1−0.2−0.8
    dnaJ3PFREUD_08760Chaperone protein DnaJ3PM<0.01<0.01<0.01−0.40.0−0.6−1.6−1.3−0.3−0.8
    hsp20PFREUD_09500Heat shock protein 20 2PM<0.010.030.03−1.3−0.6−0.3−0.1−0.1−0.61.2
    dnaJ1PFREUD_17820Chaperone protein DnaJ1PM<0.01<0.01<0.01−2.80.0−2.2−2.7−1.6−2.3−2.2
    grpE1PFREUD_17830Co-chaperone proteinPM<0.01<0.01<0.01−3.00.1−2.8−3.2−2.0−3.0−2.3
    dnaK1PFREUD_17840Chaperone proteinPM<0.01<0.01<0.01−2.90.2−3.2−2.0−2.5−4.5−1.7
    clpB 2PFREUD_17920Chaperone proteinPM<0.01<0.01<0.01−2.9−0.4−3.5−3.3−2.1−1.9−4.1
    groL2PFREUD_1847060-kDa chaperonin 2PM<0.010.370.65−2.9−3.7−3.2−3.4−3.3−4.1−3.6
    clpB 1PFREUD_19250Chaperone proteinPM<0.01<0.01<0.011.90.7−1.30.80.22.71.2

    • ↵a Locus tag for CIRM-BIA1T.

    • ↵b A, adaptation to atypical conditions; DNA, DNA metabolism; PM, protein modification and folding.

    • ↵c Values of |fold change (log2)| >1 are in boldface.

  • Table 3

    Genes encoding esterases and branched-chain amino acid-converting enzymes

    Type of enzyme and nameLocus tagaDescriptionbCategorycP valueFold change (log2) for each CIRM-BIA straind:
    TimeStrainTime × strain11812212314724829
    Esterases
        pf1861PFREUD_03560Putative carboxylic ester hydrolaseL0.18<0.010.06−0.2−0.6−0.40.20.40.2−0.6
        pf774PFREUD_04240Putative carboxylic ester hydrolaseL0.60<0.010.110.4−0.7−0.40.30.2−0.30.6
        pf279PFREUD_04340Carboxylic ester hydrolaseL0.320.010.110.3−0.6−0.60.600.10.4
        pf962PFREUD_04810Carboxylic ester hydrolaseL<0.010.050.610.50.40.50.70.20.50.2
        pf1509PFREUD_10540Putative carboxylic ester hydrolaseL0.50<0.010.030.00.50.20.3−0.70.9−0.5
        pf1758-2887PFREUD_10790-PFREUD_10800Putative carboxylic ester hydrolaseseL0.570.020.34−0.3−0.2−0.3—c0.20.5—c
        pf1637PFREUD_12910Putative carboxylic ester hydrolaseL<0.01<0.010.070.0−0.40.30.0−0.8−0.3−0.4
        pf379PFREUD_13000Putative carboxylic ester hydrolaseL<0.01<0.01<0.010.80.70.61.20.61.60.6
        pf169PFREUD_14330Carboxylic ester hydrolaseL0.130.190.13−0.1−0.2−0.10.30.10.50.1
        pf1655PFREUD_18110Carboxylic ester hydrolaseL0.59<0.010.550.30.7−0.4−0.100−0.3
        pf667PFREUD_23150Carboxylic ester hydrolaseL<0.01<0.01<0.010.1−0.10.20.90.30.5−0.2
        pf2042PFREUD_23770Putative carboxylic ester hydrolaseL<0.01<0.010.04−0.3−0.6−0.6−0.20.1−0.6−0.4
    Branched-chain amino acid transport and conversion
        livGPFREUD_10850ABC protein of branched-chain amino acid ABC transporterAA<0.01<0.010.02−1.4−0.8−2.3−3.6−2.4−3−2.1
        braEPFREUD_10860IM protein of branched-chain amino acid ABC transporterAA<0.01<0.01<0.01−0.5−0.3−0.8−1.5−1.1−1.5−0.6
        braDPFREUD_10870IM protein of branched-chain amino acid ABC transporterAA<0.01<0.01<0.01−1.0−0.4−1.3−1.9−1−2.4−0.9
        braCPFREUD_10880BP of branched-chain amino acid ABC transporterAA<0.01<0.010.01−1.0−0.5−1.2−2.9−1.6−2.2−0.6
        ydaOPFREUD_12690IM protein of branched-chain amino acid ABC transporterAA<0.01<0.010.06−2.1−2−1.6−2.4−1.9−2.4−2.2
        ilvEPFREUD_13350Branched-chain amino acid aminotransferaseAA0.450.010.10−0.20.2−0.40.3−1.50.40.4
        bkdA2PFREUD_022002-Oxoisovalerate dehydrogenase subunit betaAA<0.01<0.010.03−2.1−3.7−2.0−3.3−3.3−3.5−3.9
        bkdBPFREUD_02210Dihydrolipoyllysine residue (2-methylpropanoyl) transferaseAA<0.01<0.010.03−0.8−1.7−0.7−1.8−1.4−0.7−1.5

    • ↵a Locus tag for CIRM-BIA1T.

    • ↵b IM, integral membrane; BP, binding protein.

    • ↵c L, lipid metabolism; AA, transport and metabolism of amino acids.

    • ↵d Values of |fold change (log2)| >1 are in boldface.

    • ↵e This gene presents a frameshift in strains CIRM-BIA1T and CIRM-BIA9.

  • Table 4

    Differentially expressed genes involved in pyruvate generation and rerouting toward trehalose and glycogen synthesis

    Function and nameLocus tagaDescriptionCategorybP valueFold change (log2) for each CIRM-BIA strainc:
    TimeStrainTime × strain11812212314724829
    Generation of energy
        ppaPFREUD_23500Inorganic pyrophosphatasePh<0.01<0.01<0.012.51.81.73.32.72.82.5
    Generation of pyruvate
        aldPFREUD_00370Alanine dehydrogenaseAA<0.01<0.01<0.013.74.51.17.53.22.04.3
        sdaAPFREUD_18570l-Serine dehydrataseAA<0.01<0.01<0.011.61.30.52.01.01.90.9
        ldh2PFREUD_12840l-Lactate dehydrogenaseCH<0.010.020.011.52.01.31.31.71.80.4
    Gluconeogenesis
        ppdkPFREUD_03230Pyruvate phosphate dikinaseCH<0.01<0.01<0.011.31.2−0.61.90.90.30.6
        eno1PFREUD_17320Enolase 1CH<0.01<0.010.011.110.62.61.91.71.1
        eno2PFREUD_17250Enolase 2CH<0.01<0.010.021.21.20.70.61.31.10.7
        fba1PFREUD_19150Fructose-bisphosphate aldolase class IICH<0.01<0.010.33−0.5−1.2−0.3−1.0−0.7−0.6−1.0
        fba2PFREUD_23890Fructose-bisphosphate aldolase class ICH<0.01<0.01<0.012.52.40.53.92.31.22.2
        pgiPFREUD_04290Glucose-6-phosphate isomeraseCH<0.01<0.01<0.011.00.40.51.60.81.21.5

    • ↵a Locus tag in CIRM-BIA1T.

    • ↵b Ph, metabolism of phosphate; AA, transport and metabolism of amino acids; CH, transport and metabolism of carbohydrates.

    • ↵c Values of |fold change (log2)| >1 are in boldface.

Back to top
Download PDF
Citation Tools
Print

Alerts
Email

Related Articles

Cited By...