BRENDA - Enzyme Database
show all sequences of 1.4.3.3

Thermostabilization of porcine kidney D-amino acid oxidase by a single amino acid substitution

Bakke, M.; Setoyama, C.; Miura, R.; Kajiyama, N.; Biotechnol. Bioeng. 93, 1023-1027 (2006)

Data extracted from this reference:

Engineering
Protein Variants
Commentary
Organism
F42C
mutant retains more than 70% of activity after 1 h, while the wild-type enzyme retains only 10% activity. Optimal temperature of mutant enzyme is about 10°C higher than that of wild-type enzyme. Activity at the optimal temperature is about 20% higher than that of wild-type enzyme. KM-values for D-Ala, D-Met, D-Phe and D-Ser are 40-50% of the wild-type value
Sus scrofa
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.4
-
D-Met
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
0.9
-
D-Met
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
1.2
-
D-Ala
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
1.9
-
D-Phe
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
2.5
-
D-Ala
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
3.6
-
D-Phe
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
4.7
-
D-Ser
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
12.7
-
D-Ser
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
Organism
Organism
UniProt
Commentary
Textmining
Sus scrofa
-
-
-
Source Tissue
Source Tissue
Commentary
Organism
Textmining
kidney
-
Sus scrofa
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
D-Ala + H2O + O2
-
672703
Sus scrofa
pyruvate + NH3 + H2O2
-
-
-
?
D-Met + H2O + O2
-
672703
Sus scrofa
4-methylthio-2-oxobutanoate + NH3 + H2O2
-
-
-
?
D-Phe + H2O + O2
-
672703
Sus scrofa
phenylpyruvate + NH3 + H2O2
-
-
-
?
D-Ser + H2O + O2
-
672703
Sus scrofa
2-oxo-3-hydroxypropionate + NH3 + H2O2
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
45
50
wild-type enzyme
Sus scrofa
55
-
mutant enzyme F42C
Sus scrofa
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
60
30°C: about 50% of maximal activity, 60°C: about 40% of maximal activity, wild-type enzyme
Sus scrofa
40
70
40°C and 70°C: about 60% of maximal activity, mutant enzyme F42C
Sus scrofa
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
45
-
wild-type enzyme is unstable above
Sus scrofa
55
-
1 h, mutant enzyme F42C retains more than 70% of activity, wild-type enzyme retains only 10% activity
Sus scrofa
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
10
wild-type enzyme and mutant enzyme F42C
Sus scrofa
pH Range
pH Minimum
pH Maximum
Commentary
Organism
7.5
11
pH 7.5 and pH 11: about 40% of maximal activity
Sus scrofa
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
5.5
9
25°C, 24 h, stable, wild-type enzyme and mutant enzyme F42C
Sus scrofa
Engineering (protein specific)
Protein Variants
Commentary
Organism
F42C
mutant retains more than 70% of activity after 1 h, while the wild-type enzyme retains only 10% activity. Optimal temperature of mutant enzyme is about 10°C higher than that of wild-type enzyme. Activity at the optimal temperature is about 20% higher than that of wild-type enzyme. KM-values for D-Ala, D-Met, D-Phe and D-Ser are 40-50% of the wild-type value
Sus scrofa
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.4
-
D-Met
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
0.9
-
D-Met
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
1.2
-
D-Ala
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
1.9
-
D-Phe
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
2.5
-
D-Ala
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
3.6
-
D-Phe
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
4.7
-
D-Ser
pH 8.3, 37°C, mutant enzyme F42C
Sus scrofa
12.7
-
D-Ser
pH 8.3, 37°C, wild-type enzyme
Sus scrofa
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
kidney
-
Sus scrofa
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
D-Ala + H2O + O2
-
672703
Sus scrofa
pyruvate + NH3 + H2O2
-
-
-
?
D-Met + H2O + O2
-
672703
Sus scrofa
4-methylthio-2-oxobutanoate + NH3 + H2O2
-
-
-
?
D-Phe + H2O + O2
-
672703
Sus scrofa
phenylpyruvate + NH3 + H2O2
-
-
-
?
D-Ser + H2O + O2
-
672703
Sus scrofa
2-oxo-3-hydroxypropionate + NH3 + H2O2
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
45
50
wild-type enzyme
Sus scrofa
55
-
mutant enzyme F42C
Sus scrofa
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
60
30°C: about 50% of maximal activity, 60°C: about 40% of maximal activity, wild-type enzyme
Sus scrofa
40
70
40°C and 70°C: about 60% of maximal activity, mutant enzyme F42C
Sus scrofa
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
45
-
wild-type enzyme is unstable above
Sus scrofa
55
-
1 h, mutant enzyme F42C retains more than 70% of activity, wild-type enzyme retains only 10% activity
Sus scrofa
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
10
wild-type enzyme and mutant enzyme F42C
Sus scrofa
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
7.5
11
pH 7.5 and pH 11: about 40% of maximal activity
Sus scrofa
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
5.5
9
25°C, 24 h, stable, wild-type enzyme and mutant enzyme F42C
Sus scrofa
Other publictions for EC 1.4.3.3
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)