Jin, YiPetricevic, MarijaJohn, AlanRaich Armendáriz, Lluís AdriàJenkins, HuwPortela de Souza, LeticiaCuskin, FionaGilbert, Harry J.Rovira i Virgili, CarmeGoddard-Borger, Ethan D.Williams, Spencer J.Davies, Gideon J.2017-07-272017-07-272016-11-082374-7951https://hdl.handle.net/2445/114446The enzymatic cleavage of β-1,4-mannans is achieved by endo-β-1,4-mannanases, enzymes involved in germination of seeds and microbial hemicellulose degradation, and which have increasing industrial and consumer product applications. β-Mannanases occur in a range of families of the CAZy sequence-based glycoside hydrolase (GH) classification scheme including families 5, 26, and 113. In this work we reveal that β-mannanases of the newly described GH family 134 differ from other mannanase families in both their mechanism and tertiary structure. A representative GH family 134 endo-β-1,4-mannanase from a Streptomyces sp. displays a fold closely related to that of hen egg white lysozyme but acts with inversion of stereochemistry. A Michaelis complex with mannopentaose, and a product complex with mannotriose, reveal ligands with pyranose rings distorted in an unusual inverted chair conformation. Ab initio quantum mechanics/molecular mechanics metadynamics quantified the energetically accessible ring conformations and provided evidence in support of a 1C4 → 3H4 → 3S1 conformational itinerary along the reaction coordinate. This work, in concert with that on GH family 124 cellulases, reveals how the lysozyme fold can be co-opted to catalyze the hydrolysis of different polysaccharides in a mechanistically distinct manner8 p.application/pdfeng(c) American Chemical Society , 2016LisozimEnzimsCatàlisiLysozymeEnzymesCatalysisinfo:eu-repo/semantics/article6679802017-07-27info:eu-repo/semantics/openAccess28058278