Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic

Hasan Ufuk Celebioglu, Morten Ejby Hansen, Avishek Majumder, Carsten Købler, Yong Jun Goh, Kristian Thorsen, Bjarne Gregers Schmidt, Sarah O'Flaherty, Maher Abou Hachem, Sampo J. Lahtinen, Susanne Jacobsen, Todd R. Klaenhammer, Susanne Brix Pedersen, Kristian Mølhave, Birte Svensson

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Whole cell and surface proteomes were analyzed together with adhesive properties of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) grown on the emerging prebiotic raffinose, exemplifying a synbiotic. Adhesion of NCFM to mucin and intestinal HT-29 cells increased three-fold after culture with raffinose versus glucose, as also visualized by scanning electron microscopy. Comparative proteomics using 2D-DIGE showed 43 unique proteins to change in relative abundance in whole cell lysates from NCFM grown on raffinose compared to glucose. Furthermore, 14 unique proteins in 18 spots of the surface subproteome underwent changes identified by differential 2DE, including elongation factor G, thermostable pullulanase, and phosphate starvation inducible stress-related protein increasing in a range of +2.1 − +4.7 fold. By contrast five known moonlighting proteins decreased in relative abundance by up to −2.4 fold. Enzymes involved in raffinose catabolism were elevated in the whole cell proteome; α-galactosidase (+13.9 fold); sucrose phosphorylase (+5.4 fold) together with metabolic enzymes from the Leloir pathway for galactose utilization and the glycolysis; β-galactosidase (+5.7 fold); galactose (+2.9/+3.1 fold) and fructose (+2.8 fold) kinases. The insights at the molecular and cellular levels contributed to the understanding of the interplay of a synbiotic composed of NCFM and raffinose with the host.
    Original languageEnglish
    JournalProteomics
    Volume16
    Issue number9
    Pages (from-to)1361-1375
    Number of pages15
    ISSN1615-9853
    DOIs
    Publication statusPublished - 2016

    Keywords

    • Adhesion
    • HT-29 cells
    • Moonlighting proteins
    • Probiotics
    • Raffinose
    • Surface and whole cell proteomes

    Cite this

    Celebioglu, Hasan Ufuk ; Hansen, Morten Ejby ; Majumder, Avishek ; Købler, Carsten ; Jun Goh, Yong ; Thorsen, Kristian ; Schmidt, Bjarne Gregers ; O'Flaherty, Sarah ; Abou Hachem, Maher ; Lahtinen, Sampo J. ; Jacobsen, Susanne ; Klaenhammer, Todd R. ; Pedersen, Susanne Brix ; Mølhave, Kristian ; Svensson, Birte. / Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic. In: Proteomics. 2016 ; Vol. 16, No. 9. pp. 1361-1375.
    @article{7cc09130c0414f45bcda7207fdf4d9ae,
    title = "Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic",
    abstract = "Whole cell and surface proteomes were analyzed together with adhesive properties of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) grown on the emerging prebiotic raffinose, exemplifying a synbiotic. Adhesion of NCFM to mucin and intestinal HT-29 cells increased three-fold after culture with raffinose versus glucose, as also visualized by scanning electron microscopy. Comparative proteomics using 2D-DIGE showed 43 unique proteins to change in relative abundance in whole cell lysates from NCFM grown on raffinose compared to glucose. Furthermore, 14 unique proteins in 18 spots of the surface subproteome underwent changes identified by differential 2DE, including elongation factor G, thermostable pullulanase, and phosphate starvation inducible stress-related protein increasing in a range of +2.1 − +4.7 fold. By contrast five known moonlighting proteins decreased in relative abundance by up to −2.4 fold. Enzymes involved in raffinose catabolism were elevated in the whole cell proteome; α-galactosidase (+13.9 fold); sucrose phosphorylase (+5.4 fold) together with metabolic enzymes from the Leloir pathway for galactose utilization and the glycolysis; β-galactosidase (+5.7 fold); galactose (+2.9/+3.1 fold) and fructose (+2.8 fold) kinases. The insights at the molecular and cellular levels contributed to the understanding of the interplay of a synbiotic composed of NCFM and raffinose with the host.",
    keywords = "Adhesion, HT-29 cells, Moonlighting proteins, Probiotics, Raffinose, Surface and whole cell proteomes",
    author = "Celebioglu, {Hasan Ufuk} and Hansen, {Morten Ejby} and Avishek Majumder and Carsten K{\o}bler and {Jun Goh}, Yong and Kristian Thorsen and Schmidt, {Bjarne Gregers} and Sarah O'Flaherty and {Abou Hachem}, Maher and Lahtinen, {Sampo J.} and Susanne Jacobsen and Klaenhammer, {Todd R.} and Pedersen, {Susanne Brix} and Kristian M{\o}lhave and Birte Svensson",
    year = "2016",
    doi = "10.1002/pmic.201500212",
    language = "English",
    volume = "16",
    pages = "1361--1375",
    journal = "Proteomics",
    issn = "1615-9853",
    publisher = "Wiley - V C H Verlag GmbH & Co. KGaA",
    number = "9",

    }

    Celebioglu, HU, Hansen, ME, Majumder, A, Købler, C, Jun Goh, Y, Thorsen, K, Schmidt, BG, O'Flaherty, S, Abou Hachem, M, Lahtinen, SJ, Jacobsen, S, Klaenhammer, TR, Pedersen, SB, Mølhave, K & Svensson, B 2016, 'Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic', Proteomics, vol. 16, no. 9, pp. 1361-1375. https://doi.org/10.1002/pmic.201500212

    Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic. / Celebioglu, Hasan Ufuk; Hansen, Morten Ejby; Majumder, Avishek; Købler, Carsten; Jun Goh, Yong; Thorsen, Kristian; Schmidt, Bjarne Gregers ; O'Flaherty, Sarah; Abou Hachem, Maher ; Lahtinen, Sampo J.; Jacobsen, Susanne; Klaenhammer, Todd R.; Pedersen, Susanne Brix; Mølhave, Kristian; Svensson, Birte.

    In: Proteomics, Vol. 16, No. 9, 2016, p. 1361-1375.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic

    AU - Celebioglu, Hasan Ufuk

    AU - Hansen, Morten Ejby

    AU - Majumder, Avishek

    AU - Købler, Carsten

    AU - Jun Goh, Yong

    AU - Thorsen, Kristian

    AU - Schmidt, Bjarne Gregers

    AU - O'Flaherty, Sarah

    AU - Abou Hachem, Maher

    AU - Lahtinen, Sampo J.

    AU - Jacobsen, Susanne

    AU - Klaenhammer, Todd R.

    AU - Pedersen, Susanne Brix

    AU - Mølhave, Kristian

    AU - Svensson, Birte

    PY - 2016

    Y1 - 2016

    N2 - Whole cell and surface proteomes were analyzed together with adhesive properties of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) grown on the emerging prebiotic raffinose, exemplifying a synbiotic. Adhesion of NCFM to mucin and intestinal HT-29 cells increased three-fold after culture with raffinose versus glucose, as also visualized by scanning electron microscopy. Comparative proteomics using 2D-DIGE showed 43 unique proteins to change in relative abundance in whole cell lysates from NCFM grown on raffinose compared to glucose. Furthermore, 14 unique proteins in 18 spots of the surface subproteome underwent changes identified by differential 2DE, including elongation factor G, thermostable pullulanase, and phosphate starvation inducible stress-related protein increasing in a range of +2.1 − +4.7 fold. By contrast five known moonlighting proteins decreased in relative abundance by up to −2.4 fold. Enzymes involved in raffinose catabolism were elevated in the whole cell proteome; α-galactosidase (+13.9 fold); sucrose phosphorylase (+5.4 fold) together with metabolic enzymes from the Leloir pathway for galactose utilization and the glycolysis; β-galactosidase (+5.7 fold); galactose (+2.9/+3.1 fold) and fructose (+2.8 fold) kinases. The insights at the molecular and cellular levels contributed to the understanding of the interplay of a synbiotic composed of NCFM and raffinose with the host.

    AB - Whole cell and surface proteomes were analyzed together with adhesive properties of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) grown on the emerging prebiotic raffinose, exemplifying a synbiotic. Adhesion of NCFM to mucin and intestinal HT-29 cells increased three-fold after culture with raffinose versus glucose, as also visualized by scanning electron microscopy. Comparative proteomics using 2D-DIGE showed 43 unique proteins to change in relative abundance in whole cell lysates from NCFM grown on raffinose compared to glucose. Furthermore, 14 unique proteins in 18 spots of the surface subproteome underwent changes identified by differential 2DE, including elongation factor G, thermostable pullulanase, and phosphate starvation inducible stress-related protein increasing in a range of +2.1 − +4.7 fold. By contrast five known moonlighting proteins decreased in relative abundance by up to −2.4 fold. Enzymes involved in raffinose catabolism were elevated in the whole cell proteome; α-galactosidase (+13.9 fold); sucrose phosphorylase (+5.4 fold) together with metabolic enzymes from the Leloir pathway for galactose utilization and the glycolysis; β-galactosidase (+5.7 fold); galactose (+2.9/+3.1 fold) and fructose (+2.8 fold) kinases. The insights at the molecular and cellular levels contributed to the understanding of the interplay of a synbiotic composed of NCFM and raffinose with the host.

    KW - Adhesion

    KW - HT-29 cells

    KW - Moonlighting proteins

    KW - Probiotics

    KW - Raffinose

    KW - Surface and whole cell proteomes

    U2 - 10.1002/pmic.201500212

    DO - 10.1002/pmic.201500212

    M3 - Journal article

    VL - 16

    SP - 1361

    EP - 1375

    JO - Proteomics

    JF - Proteomics

    SN - 1615-9853

    IS - 9

    ER -