Alcoholysis Products by a GH53 Fungal Galactanase

In this study, the native activity of an arabinogalactan endo-β-1,4-galactanase from Aspergillus niger (AnGal) was evaluated under different reaction conditions, and in the presence of various acceptor molecules during the cleavage of the β-1,4-glycosidic linkage of a chromogenic compound and lupin galactan. A combination of spectrophotometric assays, mass spectrometry and chromatography techniques provided insights into the reaction mechanism of the enzyme and its use in the synthesis of galactosides and galactooligosaccharide derivatives. In reactions containing 2-nitrophenol galactopyranoside, AnGal promoted transglycosylation, generating longer galactooligosaccharide derivatives of 2-nitrophenol that have not previously been reported for GH53 enzymes. Furthermore, new alcoholysis products have been detected when AnGal acted on lupin galactan in the presence of benzyl alcohol. To the best of our knowledge, we are first to report the synthesis of galactotriose and galactotetraose derivatives formed by endo-β-1,4-galactanase alcoholysis. This work showcases the potential of utilizing galactanases in the synthesis of valuable galactosides and galactooligosaccharides, under mild conditions from sustainable biomass sources. Potential beneficial applications may be found in several industrial fields such as in the preparation of prodrugs and prebiotics.