BRENDA - Enzyme Database

Possible role of a histidine residue in the substrate specificity of yeast D-aspartate oxidase

Takahashi, S.; Shimada, K.; Nozawa, S.; Goto, M.; Abe, K.; Kera, Y.; J. Biochem. 159, 371-378 (2016)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.4.3.1
expressed in Escherichia coli BL21(DE3) cells
Vanrija humicola
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.4.3.1
H56A
the mutant exhibits no significant activity toward acidic D-amino acids; the mutant gains the ability to utilize neutral D-amino acids as substrates, such as D-methionine, D-phenylalanine and D-glutamine
Vanrija humicola
1.4.3.1
H56D
the mutant exhibits no significant activity toward acidic D-amino acids
Vanrija humicola
1.4.3.1
H56F
the mutant exhibits no significant activity toward acidic D-amino acids
Vanrija humicola
1.4.3.1
H56K
the mutant exhibits no significant activity toward acidic D-amino acids
Vanrija humicola
1.4.3.1
H56N
the mutant exhibits no significant activity toward acidic D-amino acids; the mutant gains the ability to utilize neutral D-amino acids as substrates, such as D-methionine, D-phenylalanine and D-glutamine
Vanrija humicola
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.3.1
D-malate
87.8% residual activity at 10 mM
Vanrija humicola
1.4.3.1
malonate
62.1% residual activity at 10 mM
Vanrija humicola
1.4.3.1
additional information
not inhibited by 10 mM meso-tartrate, benzoate, anthranilate, and crotonate
Vanrija humicola
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.3.1
2.5
-
D-Asp
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
8.5
-
D-Met
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
8.8
-
D-Leu
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
14.3
-
D-Phe
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
21
-
D-Gln
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
31.8
-
D-Met
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
34.8
-
D-Phe
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
39.4
-
D-Gln
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
83.4
-
D-Asn
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
106
-
D-Glu
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.4.3.1
D-Asp + H2O + O2
Vanrija humicola
the enzyme shows high activity only with D-aspartate
oxaloacetate + NH3 + H2O2
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.4.3.1
Vanrija humicola
Q75WF1
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.3.1
D-Asn + H2O + O2
substrate for mutant enzyme H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Asn + H2O + O2
low activity
742805
Vanrija humicola
oxaloacetate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Asp + H2O + O2
the enzyme shows high activity only with D-aspartate
742805
Vanrija humicola
oxaloacetate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Gln + H2O + O2
substrate for mutant enzymes H56A and H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Glu + H2O + O2
low activity
742805
Vanrija humicola
2-oxoglutarate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Leu + H2O + O2
substrate for mutant enzyme H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Met + H2O + O2
substrate for mutant enzymes H56A and H56N
742805
Vanrija humicola
4-(methylsulfonyl)-2-oxobutanoate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Phe + H2O + O2
substrate for mutant enzymes H56A and H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
additional information
no activity with D-Ala, D-Ser, D-Pro, D-Val, D-Thr, D-Ile, D-Leu, D-Gln, D-Met, D-Phe, D-Tyr, D-Trp, D-Lys, D-His, D-Arg, and Gly
742805
Vanrija humicola
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.4.3.1
?
x * 40000, SDS-PAGE
Vanrija humicola
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.4.3.1
0.7
-
D-Leu
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
2.2
-
D-Gln
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
2.6
-
D-Glu
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
3.1
-
D-Phe
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
3.9
-
D-Asn
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
4
-
D-Met
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
4.1
-
D-Phe
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
4.5
-
D-Gln
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
11.5
-
D-Met
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
74.5
-
D-Asp
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.4.3.1
expressed in Escherichia coli BL21(DE3) cells
Vanrija humicola
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.4.3.1
H56A
the mutant exhibits no significant activity toward acidic D-amino acids; the mutant gains the ability to utilize neutral D-amino acids as substrates, such as D-methionine, D-phenylalanine and D-glutamine
Vanrija humicola
1.4.3.1
H56D
the mutant exhibits no significant activity toward acidic D-amino acids
Vanrija humicola
1.4.3.1
H56F
the mutant exhibits no significant activity toward acidic D-amino acids
Vanrija humicola
1.4.3.1
H56K
the mutant exhibits no significant activity toward acidic D-amino acids
Vanrija humicola
1.4.3.1
H56N
the mutant exhibits no significant activity toward acidic D-amino acids; the mutant gains the ability to utilize neutral D-amino acids as substrates, such as D-methionine, D-phenylalanine and D-glutamine
Vanrija humicola
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.3.1
D-malate
87.8% residual activity at 10 mM
Vanrija humicola
1.4.3.1
malonate
62.1% residual activity at 10 mM
Vanrija humicola
1.4.3.1
additional information
not inhibited by 10 mM meso-tartrate, benzoate, anthranilate, and crotonate
Vanrija humicola
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.3.1
2.5
-
D-Asp
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
8.5
-
D-Met
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
8.8
-
D-Leu
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
14.3
-
D-Phe
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
21
-
D-Gln
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
31.8
-
D-Met
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
34.8
-
D-Phe
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
39.4
-
D-Gln
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
83.4
-
D-Asn
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
106
-
D-Glu
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.4.3.1
D-Asp + H2O + O2
Vanrija humicola
the enzyme shows high activity only with D-aspartate
oxaloacetate + NH3 + H2O2
-
-
?
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.3.1
D-Asn + H2O + O2
substrate for mutant enzyme H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Asn + H2O + O2
low activity
742805
Vanrija humicola
oxaloacetate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Asp + H2O + O2
the enzyme shows high activity only with D-aspartate
742805
Vanrija humicola
oxaloacetate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Gln + H2O + O2
substrate for mutant enzymes H56A and H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Glu + H2O + O2
low activity
742805
Vanrija humicola
2-oxoglutarate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Leu + H2O + O2
substrate for mutant enzyme H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Met + H2O + O2
substrate for mutant enzymes H56A and H56N
742805
Vanrija humicola
4-(methylsulfonyl)-2-oxobutanoate + NH3 + H2O2
-
-
-
?
1.4.3.1
D-Phe + H2O + O2
substrate for mutant enzymes H56A and H56N
742805
Vanrija humicola
? + NH3 + H2O2
-
-
-
?
1.4.3.1
additional information
no activity with D-Ala, D-Ser, D-Pro, D-Val, D-Thr, D-Ile, D-Leu, D-Gln, D-Met, D-Phe, D-Tyr, D-Trp, D-Lys, D-His, D-Arg, and Gly
742805
Vanrija humicola
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.4.3.1
?
x * 40000, SDS-PAGE
Vanrija humicola
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.4.3.1
0.7
-
D-Leu
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
2.2
-
D-Gln
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
2.6
-
D-Glu
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
3.1
-
D-Phe
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
3.9
-
D-Asn
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
4
-
D-Met
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
4.1
-
D-Phe
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
4.5
-
D-Gln
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
11.5
-
D-Met
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
74.5
-
D-Asp
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.4.3.1
0.024
-
D-Glu
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.047
-
D-Asn
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.055
-
D-Gln
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.074
-
D-Leu
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.119
-
D-Phe
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.125
-
D-Met
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.213
-
D-Gln
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.218
-
D-Phe
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
1.35
-
D-Met
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
29.8
-
D-Asp
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.4.3.1
0.024
-
D-Glu
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.047
-
D-Asn
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.055
-
D-Gln
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.074
-
D-Leu
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.119
-
D-Phe
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.125
-
D-Met
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.213
-
D-Gln
mutant enzyme H56N, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
0.218
-
D-Phe
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
1.35
-
D-Met
mutant enzyme H56A, at pH 7.5 and 37°C
Vanrija humicola
1.4.3.1
29.8
-
D-Asp
wild type enzyme, at pH 7.5 and 37°C
Vanrija humicola