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Application of proteomics and other molecular methods for understanding functionality of probiotic bacteria in gastrointestinal tract of mice

Lactic acid bacteria (LAB) are important starter cultures in food industry. Some representatives are also characterised as probiotics with health promoting activity on natural way for humans and animals, respectively. Recent scientific researches are focused on development of protective biofilms and innovative mucosal vaccines based on the innocuous LAB as live oral vectors. The aim of this study was definition of functional properties of two selected LAB on molecular level: Lactococucus lactis IL1403, a model microorganism in studying genetics and physiology of LAB, and probiotic bacterium Lactobacillus acidophilus M92. For this purpose, complementary molecular methods were applied, firstly proteomics. Developed proteomic methodology enabled identification of 490 proteins of insoluble L. lactis proteome. The proteome of L. lactis after the passage of this bacterium through mouse gastrointestinal tract (GIT) was investigated in order to develop application of L. lactis as live mucosal vaccine or heterologous protein carrier. Statistical analysis enabled comparison of L. lactis proteomes observed from two different environments, M17 medium and GIT of mice, respectively. Protein biomarkers responsible for adaptation of this bacterium to GIT were identified. The proteomic approach developed on L. lactis was applied for the proteome analysis of L. acidophilus M92 with the special focus on SlpA surface proteins that have protective role during transit through GIT and are responsible for adhesion to intestinal epithelial cells of mice.

bacterial adaptation; cell envelope associated proteins; gastrointestinal tract; Lactobacillus acidophilus; Lactococcus lactis; molecular methods; probiotics; proteomics; mass spectrometry; SDS-PAGE; PFGE; vaccine

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