Journal article
Insights into physiological traits of Bifidobacterium animalis subsp. lactis BB-12 through membrane proteome analysis
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
CFB - Core Flow, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
University of Southern Denmark3
Department of Systems Biology, Technical University of Denmark4
Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark5
KTH Royal Institute of Technology6
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.
Language: | English |
---|---|
Year: | 2012 |
Pages: | 1190-1200 |
ISSN: | 18767737 and 18743919 |
Types: | Journal article |
DOI: | 10.1016/j.jprot.2011.10.031 |
ORCIDs: | Svensson, Birte |