Biely, Peter3; Cziszarava, Maria3; Agger, Jane W.4; Li, Xin-Liang5; Puchart, Vladimir3; Vranska, Maria3; Eijsink, Vincent G.H.4; Westereng, Bjørge7
1 Forest, Nature and Biomass, Department of Geosciences and Natural Resource Management, Faculty of Science, Københavns Universitet2 Department of Geosciences and Natural Resource Management, Faculty of Science, Københavns Universitet3 Institute of Chemistry, Center for Glycomics, Slovak Academy of Sciences, Bratislava4 Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences5 Youtell Biotech Inc, Bothell6 Molecular Plant Fysiology, Department of Plant Biology, Faculty of Life Sciences, Københavns Universitet7 Molecular Plant Fysiology, Department of Plant Biology, Faculty of Life Sciences, Københavns Universitet
Results The combined action of GH10 xylanase and acetylxylan esterases (AcXEs) leads to formation of neutral and acidic xylooligosaccharides with a few resistant acetyl groups mainly at their non-reducing ends. We show here that these acetyl groups serve as targets for TrCE16 AcE. The most prominent target is the 3-O-acetyl group at the non-reducing terminal Xylp residues of linear neutral xylooligosaccharides or on aldouronic acids carrying MeGlcA at the non-reducing terminus. Deacetylation of the non-reducing end sugar may involve migration of acetyl groups to position 4, which also serves as substrate of the TrCE16 esterase. Conclusion Concerted action of CtGH10 xylanase, an AcXE and TrCE16 AcE resulted in close to complete deacetylation of neutral xylooligosaccharides, whereas substitution with MeGlcA prevents removal of acetyl groups from only a small fraction of the aldouronic acids. Experiments with diacetyl derivatives of methyl β-d-xylopyranoside confirmed that the best substrate of TrCE16 AcE is 3-O-acetylated Xylp residue followed by 4-O-acetylated Xylp residue with a free vicinal hydroxyl group. General significance This study shows that CE16 acetyl esterases are crucial enzymes to achieve complete deacetylation and, consequently, complete the saccharification of acetylated xylans by xylanases, which is an important task of current biotechnology.
Biochimica Et Biophysica Acta, 2014, Vol 1840, Issue 1, p. 516-525