Structural insight into the ligand specificity of a thermostable family 51 arabinofuranosidase, Araf 51, from Clostridium thermocellum

<p>The digestion of the plant cellwall requires the concerted action of a diverse repertoire of enzyme activities. An important component of these hydrolase consortia are arabinofuranosidases, which release L-arabinofuranose moieties from a range of plant structural polysaccharides. The anaerobic bacterium Clostridium thermocellum, a highly efficient plant cell wall degrader, possesses a single ?-L-arabinofuranosidase (EC 3.2.1.55), CtAraf51A, locatedin GH51 (glycoside hydrolase family 51). The crystal structure of the enzyme has been solved in native form and in ‘Michaelis’ complexes with both?-1,5-linkedarabinotriose and?-1,3 arabinoxylobiose, both forming a hexamer in the asymmetric unit. Kinetic studies reveal that CtAraf 51A, in contrast with well-characterized GH51 enzymes including the Cellvibrio japonicus enzyme [Beylot, McKie, Voragen, Doeswijk-Voragen and Gilbert (2001) Biochem. J. 358, 607–614], catalyses the hydrolysis of ?-1,5- linked arabino-oligosaccharides and the ?-1,3 arabinosyl side chain decorations of xylan with equal efficiency. The paucityof direct hydrogen bonds with the aglycone moiety and the flexible conformation adopted by Trp178, which stacks against the sugar at the +1 subsite, provide a structural explanation for the plasticity in substrate specificity displayed by the clostridial arabinofuranosidase.Taylor, E. J., Smith, N. L., Turkenburg, J. P., D'Souza, S., Gilbert, H. J. & Davies, G. J</p>