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

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