1 research outputs found
Identification and Accurate Quantitation of Biological Oligosaccharide Mixtures
Structure-specific characterization and quantitation
is often required
for effective functional studies of oligosaccharides. Inside the gut,
HMOs are preferentially bound and catabolized by the beneficial bacteria.
HMO utility by these bacteria employs structure-specific catabolism
based on a number of glycosidases. Determining the activity of these
enzymes requires accurate quantitation of a large number of structures.
In this study, we describe a method for the quantitation of human
milk oligosaccharide (HMO) structures employing LC/MS and isotopically
labeled internal standards. Data analysis was accomplished with a
newly developed software tool, LC/MS Searcher, that employs a reference
structure library to process LC/MS data yielding structural identification
with accurate quantitation. The method was used to obtain a meta-enzyme
analysis of bacteria, the simultaneous characterization of all glycosidases
employed by bacteria for the catabolism of milk oligosaccharides.
Analysis of consumed HMO structures confirmed the utility of a β-1,3-galactosidase
in <i>Bifidobacterium longum subsp. infantis</i> ATCC 15697
(<i>B. infantis</i>). In comparison, <i>Bifidobacterium
breve</i> ATCC 15700 showed significantly less HMO catabolic
activity compared to <i>B. infantis</i>