{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T16:40:16Z","timestamp":1790008816231,"version":"4.0.1"},"reference-count":47,"publisher":"International Union of Crystallography (IUCr)","issue":"11","license":[{"start":{"date-parts":[[2014,10,23]],"date-time":"2014-10-23T00:00:00Z","timestamp":1414022400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html"},{"start":{"date-parts":[[2014,10,23]],"date-time":"2014-10-23T00:00:00Z","timestamp":1414022400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html#TDM"}],"content-domain":{"domain":["iucr.org","wiley.com","iucrj.org"],"crossmark-restriction":true},"short-container-title":["Acta Crystallogr D Biol Crystallogr","Acta Crystallogr D","Acta Crystallogr D Biol Cryst","Acta Cryst D","Acta Cryst D Biol Crystallogr","Acta Cryst D Biol Cryst","Acta Cryst Sect D","Acta Cryst Sect D Biol Cryst","Acta Crystallogr Sect D","Acta Crystallogr Sect D Biol Crystallogr"],"published-print":{"date-parts":[[2014,11,1]]},"abstract":"<jats:p>\u03b2-Mannosidases are exo-acting glycoside hydrolases (GHs) that catalyse the removal of the nonreducing end \u03b2-D-mannose from manno-oligosaccharides or mannoside-substituted molecules. They play important roles in fundamental biological processes and also have potential applications in various industries. In this study, the first fungal GH family 5 \u03b2-mannosidase (<jats:italic>Rm<\/jats:italic>Man5B) from<jats:italic>Rhizomucor miehei<\/jats:italic>was functionally and structurally characterized.<jats:italic>Rm<\/jats:italic>Man5B exhibited a much higher activity against manno-oligosaccharides than against<jats:italic>p<\/jats:italic>-nitrophenyl \u03b2-D-mannopyranoside (<jats:italic>p<\/jats:italic>NPM) and had a transglycosylation activity which transferred mannosyl residues to sugars such as fructose. To investigate its substrate specificity and transglycosylation activity, crystal structures of<jats:italic>Rm<\/jats:italic>Man5B and of its inactive E202A mutant in complex with mannobiose, mannotriose and mannosyl-fructose were determined at resolutions of 1.3, 2.6, 2.0 and 2.4\u2005\u00c5, respectively. In addition, the crystal structure of<jats:italic>R. miehei<\/jats:italic>\u03b2-mannanase (<jats:italic>Rm<\/jats:italic>Man5A) was determined at a resolution of 2.3\u2005\u00c5. Both<jats:italic>Rm<\/jats:italic>Man5A and<jats:italic>Rm<\/jats:italic>Man5B adopt the (\u03b2\/\u03b1)<jats:sub>8<\/jats:sub>-barrel architecture, which is globally similar to the other members of GH family 5. However,<jats:italic>Rm<\/jats:italic>Man5B shows several differences in the loop around the active site. The extended loop between strand \u03b28 and helix \u03b18 (residues 354\u2013392) forms a `double' steric barrier to `block' the substrate-binding cleft at the end of the \u22121 subsite. Trp119, Asn260 and Glu380 in the \u03b2-mannosidase, which are involved in hydrogen-bond contacts with the \u22121 mannose, might be essential for exo catalytic activity. Moreover, the structure of RmMan5B in complex with mannosyl-fructose has provided evidence for the interactions between the \u03b2-mannosidase and D-fructofuranose. Overall, the present study not only helps in understanding the catalytic mechanism of GH family 5 \u03b2-mannosidases, but also provides a basis for further enzymatic engineering of \u03b2-mannosidases and \u03b2-mannanases.<\/jats:p>","DOI":"10.1107\/s1399004714019762","type":"journal-article","created":{"date-parts":[[2014,10,22]],"date-time":"2014-10-22T15:40:56Z","timestamp":1413992456000},"page":"2970-2982","update-policy":"https:\/\/doi.org\/10.1107\/cm_01","source":"Crossref","is-referenced-by-count":33,"title":["Structural insights into the substrate specificity and transglycosylation activity of a fungal glycoside hydrolase family 5 \u03b2-mannosidase"],"prefix":"10.1107","volume":"70","author":[{"given":"Peng","family":"Zhou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiaojuan","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongzhou","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhen","family":"Qin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengqiang","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"329","published-online":{"date-parts":[[2014,10,23]]},"reference":[{"key":"mv5108_bb1","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1107\/S0907444909052925","volume":"66","author":"Adams","year":"2010","journal-title":"Acta Cryst. 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