Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose – an emerging prebiotic |
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Authors: | Hasan Ufuk Celebioglu Morten Ejby Avishek Majumder Carsten Købler Yong Jun Goh Kristian Thorsen Bjarne Schmidt Sarah O'Flaherty Maher Abou Hachem Sampo J. Lahtinen Susanne Jacobsen Todd R. Klaenhammer Susanne Brix Kristian Mølhave Birte Svensson |
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Affiliation: | 1. Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Lyngby, Denmark;2. Department of Micro‐ and Nanotechnology, Technical University of Denmark, Lyngby, Denmark;3. Department of Food, Bioprocessing & Nutrition Sciences, North Carolina State University, Raleigh, NC, USA;4. Active Nutrition, Nutrition & Health, DuPont, Kantvik, Finland;5. Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, Lyngby, Denmark |
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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. |
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Keywords: | Adhesion HT‐29 cells Moonlighting proteins Probiotics Raffinose Surface and whole cell proteomes |
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