Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) CodonPlus/pLysS cells | Trigonopsis variabilis |
Protein Variants | Comment | Organism |
---|---|---|
F258A | the improvement of catalytic efficiency with D-Tyr, D-Phe, and D-Leu for mutant enzyme F258A is 3.66, 11.7, and 1.5fold, respectively. The mutant is inactive with D-Ala, D-Ser, D-Lys, and D-Thr, while the activity with D-Tyr, D-Leu and especially with D-Phe significantly increases | Trigonopsis variabilis |
F258S | the improvement of catalytic efficiency with D-Tyr, D-Phe, and D-Leu for mutant enzyme F258S is 1.7, 4.75, and 6.61fold, respectively. The mutant is inactive with D-Ala, D-Ser, D-Lys, and D-Thr, while the activity with D-Tyr, D-Leu and especially with D-Phe significantly increases | Trigonopsis variabilis |
F258Y | the mutant is inactive with D-Val, D-Tyr, D-Ala, D-Ser, D-Lys, and D-Thr | Trigonopsis variabilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.068 | - |
D-Phe | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.087 | - |
D-Tyr | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.134 | - |
D-Phe | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.227 | - |
D-Leu | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.37 | - |
D-Phe | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.45 | - |
D-Trp | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.45 | - |
D-Tyr | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.46 | - |
D-Trp | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.46 | - |
D-Met | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.49 | - |
D-Trp | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.78 | - |
D-Leu | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.84 | - |
D-Tyr | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
1.19 | - |
D-Leu | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
1.68 | - |
D-Phe | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
1.81 | - |
D-Leu | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
2.7 | - |
D-Trp | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
2.9 | - |
D-Met | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
3.3 | - |
D-Met | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
4.2 | - |
D-Met | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
11.1 | - |
D-Thr | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
14.4 | - |
D-Val | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
16.7 | - |
D-Ala | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
22 | - |
D-Asn | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
22.6 | - |
D-Asn | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
29 | - |
D-Asn | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
29.3 | - |
D-Lys | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
36.6 | - |
D-Ser | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
41 | - |
D-Val | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
46 | - |
D-Val | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
50 | - |
D-Asn | Km above 50 mM, mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
38000 | - |
x * 38000, SDS-PAGE | Trigonopsis variabilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-amino acid + H2O + O2 | Trigonopsis variabilis | - |
2-oxocarboxylate + NH3 + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trigonopsis variabilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
MonoQ column chromatography and Sephadex G-25 gel filtration | Trigonopsis variabilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Ala + H2O + O2 | - |
Trigonopsis variabilis | pyruvate + NH3 + H2O2 | - |
? | |
D-amino acid + H2O + O2 | - |
Trigonopsis variabilis | 2-oxocarboxylate + NH3 + H2O2 | - |
? | |
D-Asn + H2O + O2 | - |
Trigonopsis variabilis | 2-oxosuccinamate + NH3 + H2O2 | - |
? | |
D-Leu + H2O + O2 | - |
Trigonopsis variabilis | 4-methyl-2-oxopentanoic acid + NH3 + H2O2 | - |
? | |
D-Lys + H2O + O2 | - |
Trigonopsis variabilis | 6-amino-2-oxohexanoic acid + NH3 + H2O2 | - |
? | |
D-Met + H2O + O2 | best substrate | Trigonopsis variabilis | 4-methylthio-2-oxobutanoic acid + NH3 + H2O2 | - |
? | |
D-Phe + H2O + O2 | - |
Trigonopsis variabilis | phenylpyruvate + NH3 + H2O2 | - |
? | |
D-Ser + H2O + O2 | - |
Trigonopsis variabilis | 2-oxo-3-hydroxypropionic acid + NH3 + H2O2 | - |
? | |
D-Thr + H2O + O2 | - |
Trigonopsis variabilis | 2-oxo-3-hydroxybutyrate + NH3 + H2O2 | - |
? | |
D-Trp + H2O + O2 | - |
Trigonopsis variabilis | indol-3-pyruvate + NH3 + H2O2 | - |
? | |
D-Tyr + H2O + O2 | - |
Trigonopsis variabilis | 4-hydroxyphenylpyruvic acid + NH3 + H2O2 | - |
? | |
D-Val + H2O + O2 | - |
Trigonopsis variabilis | 2-oxoisovalerate + NH3 + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 38000, SDS-PAGE | Trigonopsis variabilis |
Synonyms | Comment | Organism |
---|---|---|
D-amino acid oxidase | - |
Trigonopsis variabilis |
DAAO | - |
Trigonopsis variabilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.7 | - |
D-Asn | Km less than 0.7 s-1, mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
1.75 | - |
D-Thr | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
3.54 | - |
D-Lys | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
7.7 | - |
D-Trp | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
11.5 | - |
D-Val | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
13 | - |
D-Asn | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
14.6 | - |
D-Val | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
15.9 | - |
D-Tyr | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
20.5 | - |
D-Ser | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
22.5 | - |
D-Tyr | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
27.2 | - |
D-Phe | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
29.1 | - |
D-Leu | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
31 | - |
D-Trp | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
33 | - |
D-Trp | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
42 | - |
D-Met | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
42.4 | - |
D-Trp | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
46 | - |
D-Leu | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
47.1 | - |
D-Phe | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
50 | - |
D-Asn | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
56 | - |
D-Leu | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
58.8 | - |
D-Phe | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
62.4 | - |
D-Asn | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
63 | - |
D-Met | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
63 | - |
D-Phe | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
66.6 | - |
D-Leu | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
72 | - |
D-Tyr | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
80.5 | - |
D-Met | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
85.3 | - |
D-Val | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
89 | - |
D-Met | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
108.6 | - |
D-Ala | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Trigonopsis variabilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
D-Lys | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.16 | - |
D-Thr | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.25 | - |
D-Val | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.36 | - |
D-Val | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.56 | - |
D-Ser | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
0.59 | - |
D-Asn | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
1.7 | - |
D-Asn | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
2.8 | - |
D-Asn | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
5.9 | - |
D-Val | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
6.5 | - |
D-Ala | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
12 | - |
D-Trp | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
12.7 | - |
D-Met | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
14.8 | - |
D-Asn | Km less than 14.8 mM-1 s-1, mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
17 | - |
D-Trp | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
21 | - |
D-Met | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
22 | - |
D-Met | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
25 | - |
D-Leu | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
37 | - |
D-Phe | mutant enzyme F258Y, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
37.3 | - |
D-Leu | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
50 | - |
D-Tyr | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
56 | - |
D-Leu | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
67.3 | - |
D-Trp | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
73.9 | - |
D-Phe | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
85.8 | - |
D-Tyr | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
86.5 | - |
D-Trp | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
175 | - |
D-Met | wild type enzyme, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
183 | - |
D-Tyr | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
245 | - |
D-Leu | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
351 | - |
D-Phe | mutant enzyme F258S, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis | |
865 | - |
D-Phe | mutant enzyme F258A, in 50 mM potassium phosphate buffer, pH 8.0, at 30°C | Trigonopsis variabilis |