title: Proteomic and transcriptional analysis of Lactobacillus johnsonii PF01 during bile salt exposure by iTRAQ shotgun proteomics and quantitative RT-PCR.
abstract: Lactobacillus johnsonii PF01 has been reported to be highly resistant to bile, a key property of probiotic microorganisms. Here, we examine the nature of the bile-salt tolerance of L. johnsonii PF01. Growth inhibition and surface morphology and physiology aberrations were observed after overnight exposure to bile stress. Quantitative proteomic profiles using iTRAQ-LC-MS/MS technology identified 8307 peptides from both untreated PF01 cells and those exposed to 0.1%, 0.2%, and 0.3% bile salts. Some 215 proteins exhibited changed levels in response to bile stress; of these, levels of 94 peptides increased while those of 121 decreased. These were classified into the following categories: stress responses, cell division, transcription, translation, nucleotide metabolism, carbohydrate transport and metabolism, cell wall biosynthesis, and amino acid biosynthesis, and 16 of unidentified function. Analysis of the mRNA expression of selected genes by quantitative reverse transcriptase-PCR verified the proteomic data. Both proteomic and mRNA data provided evidence for increased phosphotransferase activity and cell wall biosynthesis. In addition, three bile salt hydrolases were significantly upregulated by bile exposure. These findings provide a basis for future evaluations of the tolerance of potential probiotic strains toward the various gastrointestinal challenges, including bile stress.
authors.affiliation: Department of Animal Resources Science, Dankook University, Cheonan 330-714, Republic of Korea.
authors.forename: Ji Yoon
authors.initials: JY
authors.lastname: Lee
authors.forename: Edward Alain B
authors.initials: EA
authors.lastname: Pajarillo
authors.forename: Min Jeong
authors.initials: MJ
authors.lastname: Kim
authors.forename: Jong Pyo
authors.initials: JP
authors.lastname: Chae
authors.forename: Dae-Kyung
authors.initials: DK
authors.lastname: Kang
mesh_terms: D001647:Bile Acids and Salts / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D006801:Humans; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D007778:Lactobacillus / Q000187:drug effects / Q000201:enzymology; D040901:Proteomics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053719:Tandem Mass Spectrometry; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D001647:Bile Acids and Salts; D006867:Hydrolases