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Literature summary for 1.4.3.3 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
Search for more literature for 1.4.3.3

Protein Variants

Protein Variants Comment Organism
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
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

KM Value [mM]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Rhodotorula toruloides P80324
-
-

Purification (Commentary)

Purification (Comment) Organism
mutant enzyme M213R Rhodotorula toruloides

Source Tissue

Source Tissue Comment Organism Textmining

Storage Stability

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
DAAO
-
 Rhodotorula toruloides

Turnover Number [1/s]

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

Cofactor

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