Crystallization (Comment) | Organism |
---|---|
crystals of W290F and W290G are grown from protein as isolated, using hanging drop vapor diffusion from 1 to 2 M (NH4)2SO4 and 0.1 M sodium acetate at pH 4.0-5.0. To prepare the azide complex, wild-type galactose oxidase is dialyzed into 50 mM PIPES (pH 7.0) for 2 h, before addition of 20 mM sodium iethyl dithiocarbamate (DDC). The copper chelator DDC promotes the formation of orthorhombic crystals, which have more favorable crystal packing for substrate soaking experiments. The copper-free protein is crystallized in 13.5% PEG 8000, 200 mM calcium acetate, and 100 mM MES (pH 6.3) | Fusarium graminearum |
Protein Variants | Comment | Organism |
---|---|---|
W290F | kcat/KM for D-galactose is 49fold lower than wild-type value, kcat/Km for 2-methylene-1,3-propanediol is 1.2fold higher than wild-type value | Fusarium graminearum |
W290G | kcat/KM for D-galactose is 6370fold lower than wild-type value | Fusarium graminearum |
W290H | kcat/KM for D-galactose is 1180fold lower than wild-type value | Fusarium graminearum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
29 | - |
2-methylene-1,3-propanediol | 25°C, pH 7.0, mutant enzyme W290F | Fusarium graminearum | |
45 | - |
D-galactose | 25°C, pH 7.0, mutant enzyme W290H | Fusarium graminearum | |
57 | - |
2-methylene-1,3-propanediol | 25°C, pH 7.0, wild-type enzyme | Fusarium graminearum | |
82 | - |
D-galactose | 25°C, pH 7.0, wild-type enzyme | Fusarium graminearum | |
1686 | - |
D-galactose | 25°C, pH 7.0, mutant enzyme W290G | Fusarium graminearum | |
2950 | - |
D-galactose | 25°C, pH 7.0, mutant enzyme W290F | Fusarium graminearum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
copper | copper content: 1.2 mol of Cu per mol of wild-type enzyme, 0.96 mol of Cu per mol of mutant enzyme W290F, 1 mol Cu per mol of mutant enzyme W290G, 1.3 mol of Cu per mol of mutant enzyme W290H | Fusarium graminearum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Fusarium graminearum | P0CS93 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Fusarium graminearum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-methylene-1,3-propanediol + O2 | - |
Fusarium graminearum | ? | - |
? | |
D-galactose + O2 | - |
Fusarium graminearum | D-galacto-hexodialdose + H2O2 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.24 | - |
D-galactose | 25°C, pH 7.0, mutant enzyme W290H | Fusarium graminearum | |
1.66 | - |
D-galactose | 25°C, pH 7.0, mutant enzyme W290G | Fusarium graminearum | |
166 | - |
2-methylene-1,3-propanediol | 25°C, pH 7.0, mutant enzyme W290F | Fusarium graminearum | |
283 | - |
2-methylene-1,3-propanediol | 25°C, pH 7.0, wild-type enzyme | Fusarium graminearum | |
371 | - |
D-galactose | 25°C, pH 7.0, mutant enzyme W290F | Fusarium graminearum | |
503 | - |
D-galactose | 25°C, pH 7.0, wild-type enzyme | Fusarium graminearum |