Cloned (Comment) | Organism |
---|---|
overexpressed in Escherichia coli BL21(DE3) as a His-tagged fusion protein | Geotrichum |
Crystallization (Comment) | Organism |
---|---|
by using the hanging-drop vapor-diffusion method. X-ray crystal structure of the OXG-RCBH-substrate complex is determined to a resolution of 2.4 A. OXG-RCBH consists of two seven bladed beta-propeller domains. There is a large cleft between the two domains, and a unique loop encloses one side of the active site cleft. Substrate bound to the cleft, and its reducing end is arranged near the loop region that is believed to impart OXG-RCBH with its activity | Geotrichum |
Protein Variants | Comment | Organism |
---|---|---|
D465N | inactive OXG-RCBH is co-crystallized with a heptaxyloglucan substrate (XXXG) under conditions similar to the wild-type. There are no differences in the overall structure between the mutant and the wild-type | Geotrichum |
DELTAG375-H385 | a deltaloop mutant (deletion mutant of the loop region, Gly375His385) is constructed. The mutant shows endo-activity with altered substrate recognition. More specifically, cleavage occurs randomly instead of at specific sites, most likely due to the misalignment of the substrate within the subsite. It is supposed that the loop imparts unique substrate specificity with exo-mode hydrolysis in OXG-RCBH | Geotrichum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geotrichum | - |
sp. M128 | - |
Purification (Comment) | Organism |
---|---|
purified by using a HiTrap DEAE FF column | Geotrichum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
xyloglucan tetradecasaccharide | enzyme substrate specificity to the tetradecasaccharide, XXXGXXXG, is analyzed. Wild-type OXG-RCBH generates XXXGXX and XG due to its strict specificity | Geotrichum | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
oligoxyloglucan reducing end-specific cellobiohydrolase | - |
Geotrichum |
OXG-RCBH | - |
Geotrichum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
assay at | Geotrichum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
assay at | Geotrichum |