{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T19:14:30Z","timestamp":1775157270190,"version":"3.50.1"},"reference-count":54,"publisher":"International Union of Crystallography (IUCr)","issue":"11","license":[{"start":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T00:00:00Z","timestamp":1477612800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html"},{"start":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T00:00:00Z","timestamp":1477612800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html#TDM"}],"funder":[{"DOI":"10.13039\/501100001407","name":"Department of Biotechnology , Ministry of Science and Technology","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001407","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["iucr.org","wiley.com","iucrj.org"],"crossmark-restriction":true},"short-container-title":["Acta Crystallogr D Struct Biol","Acta Crystallogr D","Acta Cryst D","Acta Cryst D Struct Biol","Acta Cryst Sect D","Acta Cryst Sect D Struct Biol","Acta Crystallogr Sect D","Acta Crystallogr Sect D Struct Biol","Acta Crystallogr D Biol Crystallogr","Acta Cryst D Biol Cryst"],"published-print":{"date-parts":[[2016,11,1]]},"abstract":"<jats:p>Glucuronoxylan endo-\u03b2-1,4-xylanases cleave the xylan chain specifically at sites containing 4-<jats:italic>O<\/jats:italic>-methylglucuronic acid substitutions. These enzymes have recently received considerable attention owing to their importance in the cooperative hydrolysis of heteropolysaccharides. However, little is known about the hydrolysis of glucuronoxylans in extreme environments. Here, the structure of a thermostable family 30 glucuronoxylan endo-\u03b2-1,4-xylanase (<jats:italic>Ct<\/jats:italic>Xyn30A) from<jats:italic>Clostridium thermocellum<\/jats:italic>is reported.<jats:italic>Ct<\/jats:italic>Xyn30A is part of the cellulosome, a highly elaborate multi-enzyme complex secreted by the bacterium to efficiently deconstruct plant cell-wall carbohydrates.<jats:italic>Ct<\/jats:italic>Xyn30A preferably hydrolyses glucuronoxylans and displays maximum activity at pH 6.0 and 70\u00b0C. The structure of<jats:italic>Ct<\/jats:italic>Xyn30A displays a (\u03b2\/\u03b1)<jats:sub>8<\/jats:sub>TIM-barrel core with a side-associated \u03b2-sheet domain. Structural analysis of the<jats:italic>Ct<\/jats:italic>Xyn30A mutant E225A, solved in the presence of xylotetraose, revealed xylotetraose-cleavage oligosaccharides partially occupying subsites \u22123 to +2. The sugar ring at the +1 subsite is held in place by hydrophobic stacking interactions between Tyr139 and Tyr200 and hydrogen bonds to the OH group of Tyr227. Although family 30 glycoside hydrolases are retaining enzymes, the xylopyranosyl ring at the \u22121 subsite of<jats:italic>Ct<\/jats:italic>Xyn30A-E225A appears in the \u03b1-anomeric configuration. A set of residues were found to be strictly conserved in glucuronoxylan endo-\u03b2-1,4-xylanases and constitute the molecular determinants of the restricted specificity displayed by these enzymes.<jats:italic>Ct<\/jats:italic>Xyn30A is the first thermostable glucuronoxylan endo-\u03b2-1,4-xylanase described to date. This work reveals that substrate recognition by both thermophilic and mesophilic glucuronoxylan endo-\u03b2-1,4-xylanases is modulated by a conserved set of residues.<\/jats:p>","DOI":"10.1107\/s2059798316014376","type":"journal-article","created":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T16:37:36Z","timestamp":1477672656000},"page":"1162-1173","update-policy":"https:\/\/doi.org\/10.1107\/cm_01","source":"Crossref","is-referenced-by-count":13,"title":["Conservation in the mechanism of glucuronoxylan hydrolysis revealed by the structure of glucuronoxylan xylanohydrolase (<i>Ct<\/i>Xyn30A) from<i>Clostridium thermocellum<\/i>"],"prefix":"10.1107","volume":"72","author":[{"given":"Filipe","family":"Freire","sequence":"first","affiliation":[]},{"given":"Anil","family":"Verma","sequence":"additional","affiliation":[]},{"given":"Pedro","family":"Bule","sequence":"additional","affiliation":[]},{"given":"Victor D.","family":"Alves","sequence":"additional","affiliation":[]},{"given":"Carlos M. 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