Cloned (Comment) | Organism |
---|---|
expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3), induction by DL-Ala | Trigonopsis variabilis |
Protein Variants | Comment | Organism |
---|---|---|
C108D | site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108D, overview | Trigonopsis variabilis |
C108S | site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108S, overview | Trigonopsis variabilis |
R110A | site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme, but shows increased temperature denaturation. Release of FAD is the dominant path of thermal denaturation of R110A | Trigonopsis variabilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | oxidative modification of Trigonopsis variabilis D-amino acid oxidase in vivo is traceable as the conversion of Cys108 into a stable cysteine sulfinic acid, causes substantial loss of activity and thermostability of the enzyme | Trigonopsis variabilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-methionine + H2O + O2 | Trigonopsis variabilis | - |
4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? | |
D-methionine + H2O + O2 | Trigonopsis variabilis ATCC 10679 | - |
4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trigonopsis variabilis | - |
- |
- |
Trigonopsis variabilis ATCC 10679 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by ultracentrifugation, affinity and anion exchange chromatography | Trigonopsis variabilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.2 | - |
recombinant Strep-tagged wild-type enzyme, substrates are D-methionine and 6-dichloroindophenolate, pH 7.5, 37°C | Trigonopsis variabilis |
2.6 | - |
recombinant Strep-tagged mutant R101A, substrates are D-methionine and 6-dichloroindophenolate, pH 7.5, 37°C | Trigonopsis variabilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-methionine + 2,6-dichloroindophenol | - |
Trigonopsis variabilis | 4-methylsulfanyl-2-oxobutanoate + reduced 2,6-dichloroindophenol | - |
? | |
D-methionine + 2,6-dichloroindophenol | - |
Trigonopsis variabilis ATCC 10679 | 4-methylsulfanyl-2-oxobutanoate + reduced 2,6-dichloroindophenol | - |
? | |
D-methionine + H2O + O2 | - |
Trigonopsis variabilis | 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? | |
D-methionine + H2O + O2 | with 6-dichloroindophenol as electron acceptor | Trigonopsis variabilis | 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? | |
D-methionine + H2O + O2 | - |
Trigonopsis variabilis ATCC 10679 | 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? | |
D-methionine + H2O + O2 | with 6-dichloroindophenol as electron acceptor | Trigonopsis variabilis ATCC 10679 | 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | - |
Trigonopsis variabilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Trigonopsis variabilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.84 | - |
D-methionine | recombinant mutant C108S, pH 7.5, 37°C, with 6-dichloroindophenolate | Trigonopsis variabilis | |
1.5 | - |
D-methionine | recombinant wild-type enzyme, pH 7.5, 37°C, with 6-dichloroindophenolate | Trigonopsis variabilis | |
2.8 | - |
D-methionine | recombinant mutant C108D, pH 7.5, 37°C, with 6-dichloroindophenolate | Trigonopsis variabilis | |
75 | - |
D-methionine | recombinant mutant C108D, pH 7.5, 37°C, with O2 | Trigonopsis variabilis | |
110 | - |
D-methionine | recombinant mutant C108S, pH 7.5, 37°C, with O2 | Trigonopsis variabilis | |
200 | - |
D-methionine | recombinant wild-type enzyme, pH 7.5, 37°C, with O2 | Trigonopsis variabilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Trigonopsis variabilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Trigonopsis variabilis |
Organism | Comment | Expression |
---|---|---|
Trigonopsis variabilis | DL-Ala induces the enzyme | up |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
153 | - |
D-methionine | recombinant mutant C108S, pH 7.5, 37°C | Trigonopsis variabilis | |
258 | - |
D-methionine | recombinant mutant C108D, pH 7.5, 37°C | Trigonopsis variabilis | |
274 | - |
D-methionine | recombinant wild-type enzyme, pH 7.5, 37°C | Trigonopsis variabilis |