EC Number   |
Protein Variants |
Reference |
|---|
   1.1.99.B3 | C130A |
FAD binding site, FAD covalently attached |
698829 |
| 1.1.99.B3 | H70A |
FAD binding site, FAD covalently attached |
698829 |
| 1.1.99.B3 | Y310A |
larger amount of carbohydrates |
698829 |
| 1.1.99.B3 | H70A/C130A |
no activity, lack of essential FAD cofactor |
698829 |
| 1.1.99.B3 | W351F |
the mutant shows reduced kcat values for monosaccharide and oligosaccharide substrates |
-, 724616 |
| 1.1.99.B3 | Y300A |
the mutant variant of gluco-oligosaccharide oxidase (GOOX) from Sarocladium strictum shows a broader substrate range and higher H2O2 stability compared to the wild-type enzyme. The mutant Y300A exhibits up to 40times higher activity on all tested sugars except glucose, compared to wild-type. Fusion of the Y300A variant to a family 22 carbohydrate binding module from Clostridium thermocellum (CtCBM22A) nearly doubles its catalytic efficiency on glucose, while retaining significant activity on oligosaccharides. In the presence of 200 mM of H2O2, the recombinant CtCBM22A_Y300A retains 80% of activity on glucose and 100% of activity on cellobiose, the preferred substrate for this enzyme, while the wild-type enzyme retains 60% activity on D-glucose under the same conditions. GOOX variants appear to undergo a different mechanism of inactivation, as a loss of histidine instead of methionine is observed after H2O2 incubation. The addition of CtCBM22A also promotes functional binding of the fusion enzyme to xylan, facilitating its simultaneous purification and immobilization using edible oat spelt xylan, which might benefit the usage of this enzyme preparation in food and baking applications. The presence of CtCBM22A also permits enzyme binding to oat spelt xylans, facilitating simultaneous purification and immobilization of the enzyme, and ancillary fibre enrichment in potential food applications |
743824 |
| 1.1.99.B3 | E247A |
the mutation decreases the kcat and catalytic efficiency of the enzyme on oligosaccharides compared to the wild type enzyme |
-, 740204 |
| 1.1.99.B3 | Q353A |
the mutation decreases the kcat and catalytic efficiency of the enzyme on oligosaccharides compared to the wild type enzyme |
740204 |
| 1.1.99.B3 | Q353N |
the mutation decreases the kcat and catalytic efficiency of the enzyme on oligosaccharides compared to the wild type enzyme |
740204 |
| 1.1.99.B3 | Q384A |
the mutation decreases the kcat and catalytic efficiency of the enzyme on oligosaccharides compared to the wild type enzyme |
-, 740204 |
