1.4.3.3: D-amino-acid oxidase

This is an abbreviated version!
For detailed information about D-amino-acid oxidase, go to the full flat file.

Word Map on EC 1.4.3.3

Reaction

a D-amino acid
+
H2O
+
O2
=
a 2-oxo carboxylate
+
NH3
+
H2O2

Synonyms

chDAO, D-AAO, D-amino acid oxidase, D-amino acid oxidases, D-amino-acid-oxidase, D-aminoacid oxidase, DAAO, DAMOX, DAO, DAO1, DaoE, hDAAO, ophio-amino-acid oxidase, oxidase, D-amino acid, PEG-DAO, pkDAAO, RgDAAO, TvDAAO, TvDAO, LH99

ECTree

     1 Oxidoreductases
         1.4 Acting on the CH-NH2 group of donors
             1.4.3 With oxygen as acceptor
                1.4.3.3 D-amino-acid oxidase

Engineering

Engineering on EC 1.4.3.3 - D-amino-acid oxidase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G281C
G72
-
the truncation mutant shows enhanced enzyme activity
G72123-153
-
the truncation mutant shows enhanced enzyme activity
G72138-153
-
the truncation mutant shows enhanced enzyme activity
R199W
-
naturally occuring mutation, the mutation in the D-amino acid oxidase gene is associated with classical adult onset familial amyotrophic lateral sclerosis the 14.52 cM region on chromosome 12q22-23 is linked to disease. Neuronal cell lines expressing R199W DAO show decreased viability and increased ubiquitinated aggregates compared with cells expressing the wild-type protein, overview. Lentiviral-mediated expression of mutant R199W DAO in primary motor neuron cultures causes increased TUNEL labeling
D481N
G181R
G181R/D481N
-
compared to animals with only the Grin1D481N mutation, mice with both the Dao1G181R and Grin1D481N mutations display an improvement in social approach and spatial memory retention, as well as a reversal of abnormally persistent latent inhibition and a partial normalization of startle responses
D481N
-
homozygous Grin1D481N mutant mice with reduced NMDA-NR1 glycine affinity exhibit an altered anxiety-like behavior. Deficient DAO activity also reverses the anxiolytic effects of diminished NMDAR function in mice carrying both the homozygous Grin1D481N and Dao1G181R mutation, phenotypes, overview
-
G181R
-
homozygous Dao1G181R mutant mice that lack function of the D-serine catabolic enzyme DAO display an elevation in anxiety, homozygous Grin1D481N mutant mice with reduced NMDA-NR1 glycine affinity exhibit also an altered anxiety-like behavior. Deficient DAO activity also reverses the anxiolytic effects of diminished NMDAR function in mice carrying both the homozygous Grin1D481N and Dao1G181R mutation, phenotypes, overview
-
D242V/Q253R/D304V
mutant with altered substrate specificity
DELTAS308-K321
-
deletion of 14 amino acids from Ser308 to Lys321 in a surface loop (connecting beta-strands 12 and 13) transforms the enzyme from a dimeric protein into a stable monomer. The mutant enzyme is still catalytically competent and retains its binding with the FAD coenzyme. The Kd value of the apoprotein-FAD complex is 5fold higher than that of the wild-type enzyme. 1.9fold increase in Km-value for D-Ala, 1.8fold decrease in Vmax value with D-Trp
G52V
-
site-directed mutagenesis, in the mutant the reactivity of the reduced enzyme with O2 is decreased about 100fold and the turnover number about 1000fold compared to the wild-type enzyme
L118H
mutant with altered substrate specificity
M213E
M213G
M213R
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
Q144R
Q339E
Q339N
-
mutant shows lower activity on neutral and basic amino acids compared to the wild type enzyme
R285A
-
decreased activity with D-amino acids
R285A/D/K/Q
-
inactive
R285D
-
decreased activity with D-amino acids
R285K
-
decreased activity with D-amino acids
R285Q
-
decreased activity with D-amino acids
S19G/S120P/Q144R/K321M/A345V
S335G
S335H
-
mutant shows lower activity on neutral and basic amino acids compared to the wild type enzyme
S335R
-
mutant shows lower activity on neutral and basic amino acids compared to the wild type enzyme
T60A/Q144R/K152E
-
increased overall activity compared to the wild type enzyme
W243I
-
mutant ezyme with a significantly lower content of flavin cofactor. Loss of the tertiary structure elements and of the flavin cofactor occurs at lower temperatures in mutant than in the dimeric wild-type enzyme. The midpoint concentration of urea required for unfolding is significanlty lower than for the wild-type enzyme
W243Y
-
mutant enzyme retains the FAD coenzyme. Loss of the tertiary structure elements and of the flavin cofactor occurs at lower temperatures in mutant than in the dimeric wild-type enzyme
Y223F
-
mutant shows lower activity on neutral and basic amino acids compared to the wild type enzyme
Y223S
Y238F
Y238R
Y238S
E220D/Y224G
E222D/Y224G
-
decreased catalytic activity for D-Ala compared to the wild type enzyme
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 10C 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
G313A
H307L
-
Kd for FAD is 28fold higher with respect to the wild type enzyme while activity is mostly retained
R221D/Y224G
-
decreased catalytic activity for D-Ala compared to the wild type enzyme
T317A
-
decreased activity to FAD
Y224F
-
turnover numbers similar to wild-type
Y228F
C106C108-(SO2H)
-
oxidatively modified enzyme shows 75% loss of activity
C108D
-
site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108D, overview
C108S
-
site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108S, overview
D206A
-
decreased activity with D-amino acids
D206E
-
decreased activity with D-amino acids
D206G
-
no activity with D-amino acids
D206L
-
no activity with D-amino acids
D206N
-
no activity with D-amino acids
D206S
-
decreased activity with D-amino acids
F258A
-
the improvement of catalytic efficiency with D-Tyr, D-Phe, and D-Leu for mutant enzyme F258A is 3.66, 11.7, and 1.5fold, respectively. The mutant is inactive with D-Ala, D-Ser, D-Lys, and D-Thr, while the activity with D-Tyr, D-Leu and especially with D-Phe significantly increases
F258S
-
the improvement of catalytic efficiency with D-Tyr, D-Phe, and D-Leu for mutant enzyme F258S is 1.7, 4.75, and 6.61fold, respectively. The mutant is inactive with D-Ala, D-Ser, D-Lys, and D-Thr, while the activity with D-Tyr, D-Leu and especially with D-Phe significantly increases
F258Y
-
the mutant is inactive with D-Val, D-Tyr, D-Ala, D-Ser, D-Lys, and D-Thr
F54A
-
site-directed mutagenesis
F54S
-
site-directed mutagenesis
F54Y
-
site-directed mutagenesis, the mutant shows 6fold improvement in kcat,app and about 2.5fold increase in Ki of glutaryl-7-aminocephalosporanic acid, the substitution improves the catalytic activity and thermostability of mutant DAAO compared to the wild-type enzyme. Heat treatment at 55 for 60 min does not decrease the activity of F54Y. The Tyr substitution might initiate hydrogen bond formation with the amino group of CPC and facilitate deamination
H324L
-
inactive
H324N
-
decreased activity
H324Q
-
decreased activity
H324R
-
inactive
M156L
-
mutant shows increased H2O2 resistance
M209L
-
mutant shows increased H2O2 resistance
R110A
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme, but shows increased temperature denaturation. Release of FAD is the dominant path of thermal denaturation of R110A
Y223F
-
slower substrate binding than the wild-type
C108D
-
site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108D, overview
-
C108S
-
site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108S, overview
-
R110A
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme, but shows increased temperature denaturation. Release of FAD is the dominant path of thermal denaturation of R110A
-
additional information