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 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
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