A family 10 Thermoascus aurantiacus xylanase utilizes arabinose decorations of xylan as significant substrate specificity determinants

Vardakou, M; Flint, J; Christakopoulos, P; Lewis, RJ; Gilbert, HJ; Murray, JW

dc.contributor.author	Vardakou, M	en
dc.contributor.author	Flint, J	en
dc.contributor.author	Christakopoulos, P	en
dc.contributor.author	Lewis, RJ	en
dc.contributor.author	Gilbert, HJ	en
dc.contributor.author	Murray, JW	en
dc.date.accessioned	2014-03-01T01:21:43Z
dc.date.available	2014-03-01T01:21:43Z
dc.date.issued	2005	en
dc.identifier.issn	0022-2836	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/16335
dc.subject	3D structure	en
dc.subject	Xylan	en
dc.subject	Xylanase	en
dc.subject.classification	Biochemistry & Molecular Biology	en
dc.subject.other	xylan endo 1,3 beta xylosidase	en
dc.subject.other	article	en
dc.subject.other	catalysis	en
dc.subject.other	controlled study	en
dc.subject.other	crystal structure	en
dc.subject.other	enzyme activity	en
dc.subject.other	nonhuman	en
dc.subject.other	plant cell	en
dc.subject.other	priority journal	en
dc.subject.other	protein interaction	en
dc.subject.other	Thermoascus	en
dc.subject.other	Arabinose	en
dc.subject.other	Cell Wall	en
dc.subject.other	Chromatography, High Pressure Liquid	en
dc.subject.other	Crystallography, X-Ray	en
dc.subject.other	Endo-1,4-beta Xylanases	en
dc.subject.other	Eurotiales	en
dc.subject.other	Kinetics	en
dc.subject.other	Multigene Family	en
dc.subject.other	Plant Diseases	en
dc.subject.other	Protein Structure, Tertiary	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Xylans	en
dc.subject.other	Xylose	en
dc.subject.other	Thermoascus aurantiacus	en
dc.title	A family 10 Thermoascus aurantiacus xylanase utilizes arabinose decorations of xylan as significant substrate specificity determinants	en
heal.type	journalArticle	en
heal.identifier.primary	10.1016/j.jmb.2005.07.051	en
heal.identifier.secondary	http://dx.doi.org/10.1016/j.jmb.2005.07.051	en
heal.language	English	en
heal.publicationDate	2005	en
heal.abstract	Xylan, which is a key component of the plant cell wall, consists of a backbone of beta-1,4-linked xylose residues that are decorated with arabinofuranose, acetyl, 4-O-methyl D-glucuronic acid and ferulate. The backbone of xylan is hydrolysed by endo-beta 1,4-xylanases (xylanases); however, it is unclear whether the various side-chains of the polysaccharide are utilized by these enzymes as significant substrate specificity determinants. To address this question we have determined the crystal structure of a family 10 xylanase from Thermoascus aurantiacus, in complex with xylobiose containing an arabinofuranosyl-ferulate side-chain. We show that the distal glycone subsite of the enzyme makes extensive direct and indirect interactions with the arabinose side-chain, while the ferulate moiety is solvent-exposed. Consistent with the 3D structural data, the xylanase displays fourfold more activity against xylotriose in which the non-reducing moiety is linked to an arabinose side-chain, compared to the undecorated form of the oligosacchairde. These data indicate that the sugar decorations of xylans in the T. aurantiacus family 10 xylanase, rather than simply being accommodated, can be significant substrate specificity determinants. (c) 2005 Elsevier Ltd. All rights reserved.	en
heal.publisher	ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD	en
heal.journalName	Journal of Molecular Biology	en
dc.identifier.doi	10.1016/j.jmb.2005.07.051	en
dc.identifier.isi	ISI:000232187100005	en
dc.identifier.volume	352	en
dc.identifier.issue	5	en
dc.identifier.spage	1060	en
dc.identifier.epage	1067	en