title: Insights into physiological traits of Bifidobacterium animalis subsp. lactis BB-12 through membrane proteome analysis.
abstract: Bifidobacterium animalis subsp. lactis BB-12 is a widely used probiotic strain associated with a variety of health-promoting traits. There is, however, only limited knowledge available regarding the membrane proteome and the proteins involved in oligosaccharide transport in BB-12. We applied two enrichment strategies to improve the identification of membrane proteins from BB-12 cultures grown on glucose and on xylo-oligosaccharides, the latter being an emerging prebiotic substrate recently reported to be fermented by BB-12. Our approach encompassed consecutive steps of detergent- and carbonate-treatment in order to generate inside-out membrane vesicles and to interfere with binding of membrane-associated proteins to the membrane, respectively. Proteins in the enriched membrane fraction and membrane-associated fraction were digested by lysyl endopeptidase and trypsin followed by peptide sequencing by LC-ESI-Q-TOF MS/MS. Ninety of a total of 248 identified unique proteins were predicted to possess transmembrane segments (TMSs), and 56 of these have more than one TMS. Seventy-nine of the identified proteins are annotated to be involved in transport of amino acids, oligosaccharides, inorganic ions, nucleotides, phosphate or exopolysaccharides, or to belong to the F1F0-ATP-synthetase complex and the protein translocation machinery, respectively.
authors: Ofir
authors: O
authors: Gilad
authors: Karin
authors: K
authors: Hjernø
authors: Eva C
authors: EC
authors: Østerlund
authors: Abelardo
authors: A
authors: Margolles
authors: Birte
authors: B
authors: Svensson
authors: Birgitte
authors: B
authors: Stuer-Lauridsen
authors: Anders L Blom
authors: AL
authors: Møller
authors: Susanne
authors: S
authors: Jacobsen
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D001644:Bifidobacterium / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods; D003902:Detergents / Q000494:pharmacology; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009844:Oligosaccharides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D019936:Probiotics / Q000737:chemistry; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012697:Serine Endopeptidases / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014357:Trypsin / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D003902:Detergents; D009844:Oligosaccharides; D010455:Peptides; D020543:Proteome; D012697:Serine Endopeptidases; D014357:Trypsin; C036813:lysyl endopeptidase; D005947:Glucose