Effect of acid stress on protein expression and phosphorylation in Lactobacillus rhamnosus GG

• Published in: Journal of proteomics 2012-02, Vol.75 (4), p.1357-1374
• Main Authors: Koponen, Johanna, Laakso, Kati, Koskenniemi, Kerttu, Kankainen, Matti, Savijoki, Kirsi, Nyman, Tuula A., de Vos, Willem M., Tynkkynen, Soile, Kalkkinen, Nisse, Varmanen, Pekka
• Format: Article
• Language: English
• Subjects: 2-D DIGE; antigens; bacillus-subtilis; Bacteria; Bioreactors; cultured cells; Data processing; DNA microarrays; Enzymes; Food; gastrointestinal system; Gastrointestinal tract; gels; Gene expression; genes; Glycolysis; gram-positive bacteria; heat-shock; Lactic acid; lactic acid bacteria; Lactobacillus rhamnosus; Lactobacillus rhamnosus GG; lactococcus-lactis; low-ph; mass-spectrometry; Metabolism; Nucleotides; pH effects; Phosphorylation; Probiotic; probiotic bacteria; probiotics; Protein biosynthesis; protein phosphorylation; protein synthesis; proteins; proteome; proteomic analysis; proteomics; Pyruvic acid; ser/thr/tyr phosphoproteome; Stress; surface antigens; Survival; tolerance response; transcriptome; Whey
• ISSN: 1874-3919
• DOI: 10.1016/j.jprot.2011.11.009

Acidic environments encountered in food products and during gastrointestinal tract passage affect the survival of bacteria that are marketed as probiotics. In this study, the global proteome responses of the probiotic lactic acid bacterium Lactobacillus rhamnosus GG to two physiologically relevant pH conditions (pH 4.8 and pH 5.8) were studied by 2-D DIGE. The proteomics data were complemented with transcriptome analyses by whole-genome DNA microarrays. The cells were cultured in industrial-type whey medium under strictly defined bioreactor conditions. In total, 2-D DIGE revealed the pH-dependent formation of 92 protein spots. In response to lower pH conditions, the strongest up-regulation of all proteins was detected for a predicted surface antigen, LGG_02016. In addition, the acid pH was found to up-regulate the expression of F 0F 1-ATP synthase genes whereas the abundance of proteins participating in nucleotide biosynthesis and protein synthesis was significantly diminished. Moreover, the results suggest that L. rhamnosus GG modulates its pyruvate metabolism depending on the growth pH. Furthermore, a growth pH-dependent protein phosphorylation phenomenon was detected in several L. rhamnosus GG proteins with ProQ Diamond 2-DE gel staining. Proteins participating in central cellular pathways were shown to be phosphorylated, and the phosphorylation of glycolytic enzymes was found to be especially extensive. [Display omitted] ► We studied the response of probiotic L. rhamnosus GG to acid stress. ► 2-D DIGE revealed pH-dependent production in 65 different proteins. ► The proteomics data were complemented with transcriptome analyses. ► pH-dependent protein phosphorylation was detected in several L. rhamnosus GG proteins.