Open AccessA versatile and scalable strategy for glycoprofiling bifidobacterial consumption of human milk oligosaccharides
Author(s) -
LoCascio Riccardo G.,
Niñonuevo Milady R.,
Kronewitter Scott R.,
Freeman Samara L.,
German J. Bruce,
Lebrilla Carlito B.,
Mills David A.
Publication year - 2009
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/j.1751-7915.2008.00072.x
Subject(s) - prebiotic , bifidobacterium longum , oligosaccharide , biovar , biology , bifidobacterium bifidum , food science , biochemistry , chemistry , microbiology and biotechnology , bifidobacterium , fermentation , lactobacillus , gene
Summary Human milk contains approximately 200 complex oligosaccharides believed to stimulate the growth and establishment of a protective microbiota in the infant gut. The lack of scalable analytical techniques has hindered the measurement of bacterial metabolism of these and other complex prebiotic oligosaccharides. An in vitro , multi‐strain, assay capable of measuring kinetics of bacterial growth and detailed oligosaccharide consumption analysis by FTICR‐MS was developed and tested simultaneously on 12 bifidobacterial strains. For quantitative consumption, deuterated and reduced human milk oligosaccharide (HMO) standards were used. A custom software suite developed in house called Glycolyzer was used to process the large amounts of oligosaccharide mass spectra automatically with 13 C corrections based on de‐isotoping protocols. High growth on HMOs was characteristic of Bifidobacterium longum biovar infantis strains, which consumed nearly all available substrates, while other bifidobacterial strains tested, B. longum bv. longum , B. adolescentis , B. breve and B. bifidum , showed low or only moderate growth ability. Total oligosaccharide consumption ranged from a high of 87% for B. infantis JCM 7009 to only 12% for B. adolescentis ATCC 15703. A detailed analysis of consumption glycoprofiles indicated strain‐specific capabilities towards differential metabolism of milk oligosaccharides. This method overcomes previous limitations in the quantitative, multi‐strain analysis of bacterial metabolism of HMOs and represents a novel approach towards understanding bacterial consumption of complex prebiotic oligosaccharides.