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Literature summary extracted from

  • Sacchi, S.; Lorenzi, S.; Molla, G.; Pilone, M.S.; Rossetti, C.; Pollegioni, L.
    Engineering the substrate specificity of D-amino-acid oxidase (2002), J. Biol. Chem., 277, 27510-27516.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
1.4.3.3 M213R the ratio of turnover number to Km-value for D-Asp is 74fold higher than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Glu is 29.5fold higher than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Ala is 172fold lower than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Pro is 166fold lower than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Asn is 9.4fold lower than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Gln is 3.2fold higher than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Met is 56fold lower than that of the wild-type enzyme Rhodotorula toruloides
1.4.3.3 M213R/Y238R the ratio of turnover number to Km-value for D-Asp is 36.6fold higher than that of the wild-type enzyme, the ratio of turnover number to Km-value for D-Ala is 1000fold lower than that of the wild-type enzyme Rhodotorula toruloides

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.4.3.1 L-Tartrate
-
 Bos taurus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.3.1 0.2
-
 N-Methyl-D-aspartate
-
 Bos taurus
1.4.3.1 0.9
-
 D-proline
-
 Bos taurus
1.4.3.1 2.7
-
 D-Aspartate
-
 Bos taurus
1.4.3.1 8.8
-
 D-glutamate
-
 Bos taurus
1.4.3.3 0.2
-
 D-Met pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 0.8
-
 D-Ala pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 1.5
-
 D-Met pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 2
-
 D-Asp pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 3.8
-
 D-Gln pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 4.6
-
 D-Gln pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 11
-
 D-Asn pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 14.4
-
 D-Asn pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 17.8
-
 D-Ala pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 18
-
 N-Methyl-D-aspartate pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 18
-
 D-Asp pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 21.5
-
 D-Pro pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 28
-
 D-Asp pH 8.5, 25°C, mutant enzyme M213R/Y238R Rhodotorula toruloides
1.4.3.3 33
-
 D-Glu pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 45.8
-
 D-Ala pH 8.5, 25°C, mutant enzyme M213R/Y238R Rhodotorula toruloides
1.4.3.3 77.3
-
 D-Glu pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 1280
-
 D-Pro pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.3.3 40000
-
 2 * 40000, mutant enzyme M213R and wilde-type enzyme, SDS-PAGE Rhodotorula toruloides

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.4.3.1 D-aspartate + H2O + O2 Bos taurus
-
 oxaloacetate + NH3 + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.1 Bos taurus
-
-
-
1.4.3.3 Rhodotorula toruloides P80324
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.3 mutant enzyme M213R Rhodotorula toruloides

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.4.3.1 kidney
-
 Bos taurus
-

Storage Stability

EC Number Storage Stability Organism
1.4.3.3 -20°C, wild-type and mutant enzyme M213R are stable for several months Rhodotorula toruloides

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.1 D-aspartate + H2O + O2
-
 Bos taurus oxaloacetate + NH3 + H2O2
-
 ?
1.4.3.1 D-glutamate + H2O + O2
-
 Bos taurus 2-oxoglutarate + NH3 + H2O2
-
 ?
1.4.3.1 D-proline + H2O + O2
-
 Bos taurus 2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.1 N-methyl-D-aspartate + H2O + O2
-
 Bos taurus oxaloacetate + CH3NH2 + H2O2
-
 ?
1.4.3.3 D-Ala + H2O + O2
-
 Rhodotorula toruloides pyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-Asn + H2O + O2
-
 Rhodotorula toruloides 2-oxosuccinamate + NH3 + H2O2
-
 ?
1.4.3.3 D-Asp + H2O + O2
-
 Rhodotorula toruloides oxaloacetate + NH3 + H2O2
-
 ?
1.4.3.3 D-Glu + H2O + O2
-
 Rhodotorula toruloides alpha-ketoglutarate + NH3 + H2O2
-
 ?
1.4.3.3 D-glutamine + H2O + O2
-
 Rhodotorula toruloides 2-oxoglutaramate + NH3 + H2O2
-
 ?
1.4.3.3 D-Met + H2O + O2
-
 Rhodotorula toruloides 4-methylthio-2-oxobutanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-Pro + H2O + O2
-
 Rhodotorula toruloides 2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 N-methyl-D-aspartate + H2O + O2 wild-type enzyme shows no activity, mutant enzyme M213R is active Rhodotorula toruloides ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.3.3 dimer 2 * 40000, mutant enzyme M213R and wilde-type enzyme, SDS-PAGE Rhodotorula toruloides

Synonyms

EC Number Synonyms Comment Organism
1.4.3.1 DASPO
-
 Bos taurus
1.4.3.3 DAAO
-
 Rhodotorula toruloides

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.3.3 29
-
 D-Asp pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 60
-
 D-Glu pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 79
-
 N-Methyl-D-aspartate pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 106
-
 D-Asn pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 235
-
 D-Asp pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 278
-
 D-Ala pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 290
-
 D-Gln pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 630
-
 D-Ala pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 680
-
 D-Met pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 975
-
 D-Glu pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 1135
-
 D-Gln pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 1300
-
 D-Asn pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 1615
-
 D-Pro pH 8.5, 25°C, mutant enzyme M213R Rhodotorula toruloides
1.4.3.3 1665
-
 D-Asp pH 8.5, 25°C, mutant enzyme M213R/Y238R Rhodotorula toruloides
1.4.3.3 4600
-
 D-Met pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 4640
-
 D-Pro pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides
1.4.3.3 5000
-
 D-Ala pH 8.5, 25°C, wild-type enzyme Rhodotorula toruloides

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.3 flavin the mutant enzyme M213R has a more polar microenvironment surrounding the flavin coenzyme Rhodotorula toruloides