MALDI-TOF-MS analysis of hydrolysis products of beechwood and birchwood xylans catalyzed by xylanase from Bacillus subtilis.

Abstract

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to analyze the polymerization distribution of xylo-oligosaccharides from birchwood and beechwood xylans following hydrolysis by the xylanase (XynA) from Bacillus subtilis, which was obtained by recombinant expression and purified by Ni2+ affinity column chromatography. The hydrolysis products of birchwood and beechwood xylans were analyzed by thin layer chromatography (TLC) and MALDI-TOF-MS. In the birchwood hydrolysate, the main neutral xylo-oligosaccharides were xylobiose (X2 ) and xylotriose (X3 ), and the acidic xylo-oligosaccharides (polymerization degree 4-12) were attached via a single methyl-glucuronic acid sidechain (MeG). Both X2 and X3 were identified in the beechwood xylan hydrolysate and acidic xylo-oligosaccharides (polymerization degree 4-16), which were structurally similar to those in the birchwood xylan hydrolysate. Therefore, the recombinant xylanase, XynA, has the potential to produce xylobiose (X2 ) and xylotriose (X3 ) as well as acidic xylo-oligosaccharides (MeGXn ) that would be applied in food industry. PRACTICAL APPLICATIONS: Xylo-oligosaccharides are a novel functional food additive, which has great potential in improving the quality of food. Xylo-oligosaccharides with methyl-glucuronic acid sidechains (MeG) are acidic xylo-oligosaccharides (MeGXn), which can be applied in the preparation of drugs for the treatment of cystitis and mucopolysaccharidosis. In this study, xylanase XynA was first obtained by gene cloning and expression and then used to hydrolyze the birchwood and beechwood xylans. The polymerization distribution of xylo-oligosaccharides generated during the enzymatic digestion was then determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Through enzyme hydrolysis, we are able to produce xylobiose and xylotriose for food additives.