TY - JOUR

T1 - Insights into physiological traits of Bifidobacterium animalis subsp. lactis BB-12 through membrane proteome analysis

A1 - Gilad,Ofir

A1 - Hjernø,Karin

A1 - Østerlund,Eva Christina

A1 - Margolles,Abelardo

A1 - Svensson,Birte

A1 - Stuer-Lauridsen,Birgitte

A1 - Møller,Anders Laurell Blom

A1 - Jacobsen,Susanne

AU - Gilad,Ofir

AU - Hjernø,Karin

AU - Østerlund,Eva Christina

AU - Margolles,Abelardo

AU - Svensson,Birte

AU - Stuer-Lauridsen,Birgitte

AU - Møller,Anders Laurell Blom

AU - Jacobsen,Susanne

PB - Elsevier BV

PY - 2012

Y1 - 2012

N2 - 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.

AB - 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.

KW - Bifidobacterium

KW - LC–MS

KW - Membrane proteomics

KW - Probiotics

KW - Transporters

U2 - 10.1016/j.jprot.2011.10.031

DO - 10.1016/j.jprot.2011.10.031

JO - Journal of Proteomics

JF - Journal of Proteomics

SN - 1874-3919

IS - 4

VL - 75

SP - 1190

EP - 1200

ER -