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.
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Hasan Ufuk
authors.initials: HU
authors.lastname: Celebioglu
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Morten
authors.initials: M
authors.lastname: Ejby
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Avishek
authors.initials: A
authors.lastname: Majumder
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, Lyngby, Denmark.
authors.forename: Carsten
authors.initials: C
authors.lastname: Købler
authors.affiliation: Department of Food, Bioprocessing & Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Yong Jun
authors.initials: YJ
authors.lastname: Goh
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Kristian
authors.initials: K
authors.lastname: Thorsen
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Bjarne
authors.initials: B
authors.lastname: Schmidt
authors.affiliation: Department of Food, Bioprocessing & Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: O'Flaherty
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Maher
authors.initials: M
authors.lastname: Abou Hachem
authors.affiliation: Active Nutrition, Nutrition & Health, DuPont, Kantvik, Finland.
authors.forename: Sampo J
authors.initials: SJ
authors.lastname: Lahtinen
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Susanne
authors.initials: S
authors.lastname: Jacobsen
authors.affiliation: Department of Food, Bioprocessing & Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Todd R
authors.initials: TR
authors.lastname: Klaenhammer
authors.affiliation: Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, Søltofts Plads, Lyngby, Denmark.
authors.forename: Susanne
authors.initials: S
authors.lastname: Brix
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, Lyngby, Denmark.
authors.forename: Kristian
authors.initials: K
authors.lastname: Mølhave
authors.affiliation: Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Elektrovej, Lyngby, Denmark.
authors.forename: Birte
authors.initials: B
authors.lastname: Svensson
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D063990:Gene Ontology; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D007779:Lactobacillus acidophilus / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D058977:Molecular Sequence Annotation; D020653:Peptide Elongation Factor G / Q000235:genetics / Q000378:metabolism; D056692:Prebiotics; D019936:Probiotics / Q000378:metabolism*; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D011887:Raffinose / Q000494:pharmacology*; D013194:Staining and Labeling; D000519:alpha-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D020653:Peptide Elongation Factor G; D056692:Prebiotics; D020543:Proteome; D005964:Glucosyltransferases; C025901:sucrose phosphorylase; D006026:Glycoside Hydrolases; D000519:alpha-Galactosidase; C100181:pullulanase; D011887:Raffinose; D005690:Galactose
keywords: Adhesion; HT-29 cells; Moonlighting proteins; Probiotics; Raffinose; Surface and whole cell proteomes