Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant DDOs in Escherichia coli strain Rosetta (DE3) | Vanrija humicola |
Protein Variants | Comment | Organism |
---|---|---|
R243A | site-directed mutagenesis, inactive mutant | Vanrija humicola |
R243D | site-directed mutagenesis, the mutant exhibits different spectral features of bound flavin from that of the wild-type enzyme. R243D exhibits approximately 2fold higher DDO activity than that of the wild-type with a high substrate specificity to D-aspartate. Kinetic analysis shows that R243D mutant has a 2.5fold lower Km and a 1.6fold higher kcat for D-aspartate compared to the wild-type enzyme, resulting in approximately 4fold higher catalytic efficiency | Vanrija humicola |
R243E | site-directed mutagenesis, the mutant exhibits different spectral features of bound flavin from that of the wild-type enzyme and displays lower DDO activity. The mutant also oxidizes the substrates of DAO, D-Ala and D-Arg, albeit with very low activities | Vanrija humicola |
R243K | site-directed mutagenesis, the mutant exhibits different spectral features of bound flavin from that of the wild-type enzyme and displays lower DDO activity | Vanrija humicola |
R243M | site-directed mutagenesis, the mutant exhibits different spectral features of bound flavin from that of the wild-type enzyme and displays lower DDO activity. The mutant also oxidizes the substrates of DAO, D-Ala and D-Arg, albeit with very low activities | Vanrija humicola |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes, overview | Vanrija humicola | |
1 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243D | Vanrija humicola | |
2 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243E | Vanrija humicola | |
2.5 | - |
D-Asp | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
2.6 | - |
D-Glu | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
3.8 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243M | Vanrija humicola | |
10.3 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243K | Vanrija humicola | |
13.4 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243A | Vanrija humicola | |
15.7 | - |
N-methyl-D-Asp | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
27.9 | - |
N-methyl-D-Asp | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
106 | - |
D-Glu | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Asp + H2O + O2 | Vanrija humicola | - |
oxaloacetate + NH3 + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vanrija humicola | Q75WF1 | i.e. Asterotremella humicola | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant DDOs from Escherichia coli strain Rosetta (DE3) by metal affinity chromatography and gel filtration | Vanrija humicola |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
mutant activities with D-Ala and D-Arg, overview | Vanrija humicola |
0.04 | - |
pH 7.0, 20°C, purified recombinant mutant R243E, substrate N-methyl-D-Asp | Vanrija humicola |
0.04 | - |
pH 7.0, 20°C, purified recombinant mutant R243K, substrate N-methyl-D-Asp | Vanrija humicola |
0.09 | - |
pH 7.0, 20°C, purified recombinant mutant R243A, substrate D-Asp | Vanrija humicola |
0.5 | - |
pH 7.0, 20°C, purified recombinant mutant R243K, substrate D-Glu | Vanrija humicola |
1 | - |
pH 7.0, 20°C, purified recombinant mutant R243E, substrate D-Glu | Vanrija humicola |
1.2 | - |
pH 7.0, 20°C, purified recombinant mutant R243D, substrate N-methyl-D-Asp | Vanrija humicola |
1.5 | - |
pH 7.0, 20°C, purified recombinant mutant R243K, substrate D-Asp | Vanrija humicola |
1.6 | - |
pH 7.0, 20°C, purified recombinant wild-type enzyme, substrate D-Glu | Vanrija humicola |
1.9 | - |
pH 7.0, 20°C, purified recombinant mutant R243E, substrate D-Asp | Vanrija humicola |
9.2 | - |
pH 7.0, 20°C, purified recombinant wild-type enzyme, substrate N-methyl-D-Asp | Vanrija humicola |
13.1 | - |
pH 7.0, 20°C, purified recombinant mutant R243M, substrate D-Glu | Vanrija humicola |
18.5 | - |
pH 7.0, 20°C, purified recombinant mutant R243D, substrate D-Glu | Vanrija humicola |
86 | - |
pH 7.0, 20°C, purified recombinant mutant R243M, substrate D-Asp | Vanrija humicola |
93.7 | - |
pH 7.0, 20°C, purified recombinant wild-type enzyme, substrate D-Asp | Vanrija humicola |
181 | - |
pH 7.0, 20°C, purified recombinant mutant R243D, substrate D-Asp | Vanrija humicola |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Asp + H2O + O2 | - |
Vanrija humicola | oxaloacetate + NH3 + H2O2 | - |
? | |
D-Asp + H2O + O2 | arginine residue 243 is possibly involved in the substrate recognition, active site model, overview | Vanrija humicola | oxaloacetate + NH3 + H2O2 | - |
? | |
D-Glu + H2O + O2 | - |
Vanrija humicola | 2-oxoglutarate + NH3 + H2O2 | - |
? | |
additional information | substrate specificities of wild-type and mutants DDOs, overview | Vanrija humicola | ? | - |
? | |
N-methyl-D-Asp + H2O + O2 | - |
Vanrija humicola | oxaloacetate + methylamine + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional model of ChDDO, comparison with the crystal structure of a yeast Rhodotorula gracilis D-amino acid oxidase, EC 1.4.3.3 | Vanrija humicola |
Synonyms | Comment | Organism |
---|---|---|
DDO | - |
Vanrija humicola |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | - |
assay at | Vanrija humicola |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243E | Vanrija humicola | |
1.4 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243K | Vanrija humicola | |
62.6 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243M | Vanrija humicola | |
74.5 | - |
D-Asp | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
121 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243D | Vanrija humicola |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Vanrija humicola |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
24 | - |
D-Glu | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
136 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243K | Vanrija humicola | |
562 | - |
N-methyl-D-Asp | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
650 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243E | Vanrija humicola | |
16500 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243M | Vanrija humicola | |
29800 | - |
D-Asp | pH 7.0, 20°C, recombinant wild-type enzyme | Vanrija humicola | |
121000 | - |
D-Asp | pH 7.0, 20°C, recombinant mutant R243D | Vanrija humicola |