Monitoring the hydrolysis and transglycosylation activity of alpha-glucosidase from Aspergillus niger by nuclear magnetic resonance spectroscopy and mass spectrometry.

Alpha-glucosidase from Aspergillus niger is an enzyme that catalyzes hydrolysis of alpha-1,4 linkages and transglucosylation to form alpha-1,6 linkages. In this study, an analytical method of oligosaccharides by nuclear magnetic resonance (NMR) was used to provide quantitative estimation of the fractions of each sugar unit and was applied to characterize the alpha-glucosidase reaction. Our data indicated that alpha-glucosidase reacts with the nonreducing end of oligosaccharides to form an alpha-1,6 linkage, and then a sugar unit with two alpha-1,6 linkages is gradually produced.

[NMR analysis of a contaminant in heparin].

Recently, it has been reported that certain lots of heparin are associated with an acute, rapid onset of serious side effects indicative of allergic reaction, and (1)H NMR is one of the convenience but strong analytical methods to identify a contaminant in heparin. However, an NMR signal from the contaminant in some cases is overlapped with a satellite peak from heparin, leading a misunderstanding of the presence of the contaminant. Here, we show the satellite peak observed close to the NMR signal of the contaminant, and recommend the (13)C decoupling NMR to discriminate the satellite peak from the contaminant.