The C-terminally truncated mutant R484H β-galactosidase enzyme from Bacillus circulans ATCC 31382 synthesizes oligosaccharides from lactulose (OsLu). OsLu may act as prebiotic compounds and are potentially resistant to gut digestion.
In total nine oligosaccharide structures were identified using HPAEC-PAD chromatography, Maldi-ToF-MS, and NMR analysis. Compared with previous studies, six of the characterized structures are novel compounds (i.e. 1, 2a, 5, 6, 7a and 7b). The products of the R484H mutant enzyme have a preference for introducing β(1→4) and β(1→3) linkages. The trisaccharide β-D-Galp-(1→3)β-D-Galp-(1→4)-D-Fru is major available in the compound mixture. OsLU structures 1-5 were found in both the wild type and R484H mutant. The tetrasaccharides compounds 6 and 7 were only synthesized by the mutant enzyme.
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