Proteomic analysis of Bifidobacterium longum subsp. infantis reveals the metabolic insight on consumption of prebiotics and host glycans

• Published in: PloS one Vol. 8; no. 2; p. e57535
• Main Authors: Kim, Jae-Han, An, Hyun Joo, Garrido, Daniel, German, J Bruce, Lebrilla, Carlito B, Mills, David A
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 26.02.2013; Public Library of Science (PLoS)
• Subjects: Baby foods; Bacteria; Bacterial Proteins - metabolism; Bifidobacterium - metabolism; Bifidobacterium longum; Binding sites; Biology; Breast milk; Carbohydrate Metabolism; Carbon; Catabolism; Cell walls; Chromatography, High Pressure Liquid; Enzymes; Epimerase; Food science; Fourier transforms; Fructans; Fructooligosaccharides; Galactokinase; Galactooligosaccharides; Galactose; Glucose; Glucose metabolism; Glycoconjugates; Infants; Intestinal microflora; Intestine; Lactobacillus; Lactose; Localization; Lymphocytes B; Mass spectrometry; Metabolism; Microbiota; Milk; Mucin; N-Acetylglucosamine; Nutrition; Oligosaccharides; Phosphates; Phosphoglucomutase; Polysaccharides; Polysaccharides - metabolism; Prebiotics; Proteins; Proteomics; Scientific imaging; Substrates; Sucrose; Sugar; Tandem Mass Spectrometry; Transport; United States > US; California
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0057535

Bifidobacterium longum subsp. infantis is a common member of the intestinal microbiota in breast-fed infants and capable of metabolizing human milk oligosaccharides (HMO). To investigate the bacterial response to different prebiotics, we analyzed both cell wall associated and whole cell proteins in B. infantis. Proteins were identified by LC-MS/MS followed by comparative proteomics to deduce the protein localization within the cell. Enzymes involved in the metabolism of lactose, glucose, galactooligosaccharides, fructooligosaccharides and HMO were constitutively expressed exhibiting less than two-fold change regardless of the sugar used. In contrast, enzymes in N-Acetylglucosamine and sucrose catabolism were induced by HMO and fructans, respectively. Galactose-metabolizing enzymes phosphoglucomutase, UDP-glucose 4-epimerase and UTP glucose-1-P uridylytransferase were expressed constitutively, while galactokinase and galactose-1-phosphate uridylyltransferase, increased their expression three fold when HMO and lactose were used as substrates for cell growth. Cell wall-associated proteomics also revealed ATP-dependent sugar transport systems associated with consumption of different prebiotics. In addition, the expression of 16 glycosyl hydrolases revealed the complete metabolic route for each substrate. Mucin, which possesses O-glycans that are structurally similar to HMO did not induced the expression of transport proteins, hydrolysis or sugar metabolic pathway indicating B. infantis do not utilize these glycoconjugates.