Summary
Previous studies have demonstrated that endoglucanase is required for cellulose biosynthes is both in bacteria and plants. However, it has yet to be elucidated how the endoglucanases function in the mechanism of cellulose biosynthesis. Here we describe the crystal structure of the cellulose biosynthesis-related endo-β-1,4-glucanase (CMCax; EC 3.2.1.4) from the cellulose-producing Gram-negative bacterium, Acetobacter xylinum (=Gluconacetobacter xylinus), determined at 1.65 Å resolution, respectively. The results showed resolution. CMCax falls into the glycoside hydrolase family 8 (GH-8), and the structure showed that the overall fold of the CMCax is similar to those of other glycoside hydrolases belonging to GH-8. Structure comparison with Clostridium thermocellum CelA, the best characterized GH-8 endoglucanase, revealed that sugar recognition sub-site +3 is completely missing in CMCax. The absence of the sub-site +3 leads to significant broadness of the cleft at the cellooligosaccharide reducing-end side. CMCax is known to be a secreted enzyme and is present in the culture medium. However, electron microscopic analysis using immunostaining clearly demonstrated that a portion of CMCax is localized to the cell surface, suggesting a link with other known membrane-anchored endoglucanases that are required for cellulose biosynthesis.
References