1. Mode of action of acetylxylan esterases on acetyl glucuronoxylan and acetylated oligosaccharides generated by a GH10 endoxylanase.
- Author
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Biely P, Cziszárová M, Uhliariková I, Agger JW, Li XL, Eijsink VG, and Westereng B
- Subjects
- Acetylation, Acetylesterase metabolism, Biomass, Carbohydrate Sequence, Endo-1,4-beta Xylanases metabolism, Esterases chemistry, Esterases metabolism, Glucuronates chemistry, Glucuronates metabolism, Molecular Sequence Data, Oligosaccharides metabolism, Plant Proteins metabolism, Polysaccharides chemistry, Polysaccharides metabolism, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Xylans chemistry, Xylans metabolism, Acetylesterase chemistry, Endo-1,4-beta Xylanases chemistry, Oligosaccharides chemistry, Plant Proteins chemistry
- Abstract
Background: Substitutions on the xylan main chain are widely accepted to limit plant cell wall degradability and acetylations are considered as one of the most important obstacles. Hence, understanding the modes of action of a range of acetylxylan esterases (AcXEs) is of ample importance not only to increase the understanding of the enzymology of plant decay/bioremediation but also to enable efficient bioconversion of plant biomass., Methods: In this study, the modes of action of acetylxylan esterases (AcXEs) belonging to carbohydrate esterase (CE) families 1, 4, 5 and 6 on xylooligosaccharides generated from hardwood acetyl glucuronoxylan were compared using MALDI ToF MS. Supporting data were obtained by following enzymatic deacetylation by (1)H NMR spectroscopy., Conclusions: None of the used enzymes were capable of complete deacetylation, except from linear xylooligosaccharides which were completely deacetylated by some of the esterases in the presence of endoxylanase. A clear difference was observed between the performance of the serine-type esterases of CE families 1, 5 and 6, and the aspartate-metalloesterases of family CE4. The difference is mainly due to the inability of CE4 AcXEs to catalyze deacetylation of 2,3-di-O-acetylated xylopyranosyl residues. Complete deacetylation of a hardwood acetyl glucuronoxylan requires additional deacetylating enzyme(s)., General Significance: The results contribute to the understanding of microbial degradation of plant biomass and outline the way to achieve complete saccharification of plant hemicelluloses which did not undergo alkaline pretreatment., (© 2013.)
- Published
- 2013
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