Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain BL21(DE3)pLysS | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
H244F | shows increased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD | Bacillus subtilis |
H244N | shows increased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD | Bacillus subtilis |
H244Q | shows increased kcat value compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD | Bacillus subtilis |
M261H | shows decreased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD | Bacillus subtilis |
M261Y | shows decreased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.6 | - |
glycine | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
0.7 | - |
glycine | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
0.7 | - |
sarcosine | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
0.8 | - |
glycine | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
1.4 | - |
sarcosine | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
1.9 | - |
glycine | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
2.1 | - |
sarcosine | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
2.8 | - |
sarcosine | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
4 | 10 | D-alanine | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
11 | - |
sarcosine | apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
11 | - |
sarcosine | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
17 | - |
D-proline | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
18 | - |
glycine | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
26 | - |
D-proline | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
26 | - |
D-proline | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
27 | - |
D-proline | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
28 | - |
glycine | apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
280 | - |
D-proline | apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
285 | - |
D-proline | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
350 | - |
D-alanine | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
420 | - |
D-alanine | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
490 | - |
D-alanine | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
530 | - |
D-alanine | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
2000 | - |
D-alanine | Km above 2000 mM, apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
HiTrap chelating affinity chromatography | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-alanine + H2O + O2 | - |
Bacillus subtilis | pyruvate + NH3 + H2O2 | - |
? | |
D-proline + H2O + O2 | - |
Bacillus subtilis | ? | - |
? | |
glycine + H2O + O2 | - |
Bacillus subtilis | glyoxylate + NH3 + H2O2 | - |
? | |
sarcosine + H2O + O2 | - |
Bacillus subtilis | glyoxylate + methylamine + H2O2 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
D-proline | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
0.3 | - |
glycine | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
0.33 | - |
D-alanine | kcat above 0.33 1/s, apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
0.38 | - |
D-proline | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
0.47 | - |
D-alanine | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
0.47 | - |
sarcosine | apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 | Bacillus subtilis | |
0.5 | - |
D-proline | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
0.55 | - |
glycine | apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
0.6 | - |
glycine | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
0.6 | - |
sarcosine | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
0.62 | - |
D-proline | apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
0.73 | - |
sarcosine | apparent value for mutant enzyme M261H, at 25°C and pH 8.5 | Bacillus subtilis | |
0.75 | - |
D-alanine | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
0.78 | - |
glycine | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
0.92 | - |
D-alanine | apparent value for wild type enzyme, at 25°C and pH 8.5 | Bacillus subtilis | |
1.08 | - |
sarcosine | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
1.08 | - |
D-alanine | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
1.12 | - |
sarcosine | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
1.2 | - |
glycine | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
1.23 | - |
D-proline | apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 | Bacillus subtilis | |
1.3 | - |
glycine | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
1.33 | - |
sarcosine | apparent value for mutant enzyme H244N, at 25°C and pH 8.5 | Bacillus subtilis | |
1.35 | - |
D-proline | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis | |
1.5 | - |
D-alanine | apparent value for mutant enzyme H244F, at 25°C and pH 8.5 | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | contains one molecule of non-covalently bound FAD per subunit | Bacillus subtilis |