Information on EC 1.13.12.15 - 3,4-dihydroxyphenylalanine oxidative deaminase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.13.12.15
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RECOMMENDED NAME
GeneOntology No.
3,4-dihydroxyphenylalanine oxidative deaminase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2 L-dopa + O2 = 2 3,4-dihydroxyphenylpyruvate + 2 NH3
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Deamination
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SYSTEMATIC NAME
IUBMB Comments
3,4-dihydroxy-L-phenylalanine:oxygen oxidoreductase (deaminating)
This enzyme is one of the three enzymes involved in L-dopa (3,4-dihydroxy-L-phenylalanine) catabolism in the non-oxygenic phototrophic bacterium Rubrivivax benzoatilyticus OU5 (and not Rhodobacter sphaeroides OU5 as had been thought [1]), the other two being EC 4.3.1.22 (dihydroxyphenylalanine reductive deaminase) and EC 2.6.1.49 (3,4-dihydroxyphenylalanine transaminase). In addition to L-dopa, the enzyme can also use L-tyrosine, L-phenylalanine, L-tryptophan and glutamate as substrate, but more slowly. The enzyme is inhibited by NADH and 2-oxoglutarate.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain OU5
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Manually annotated by BRENDA team
strain OU5
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxy-L-phenylalanine + O2
3,4-dihydroxyphenylpyruvate + NH3
show the reaction diagram
aromatic amine + 1/2 O2
aromatic aldehyde + NH3 + H2O
show the reaction diagram
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oxidative deamination, unusual oxygen-consuming reaction catalyzed by the enzyme toward aromatic amines (serotonin, dopamine, and alpha-methyldopamine) and D-tryptophan methyl ester
production in equivalent amounts depending on the nature of the substrate, and ammonia with concomitant O2 consumption in a 1:2 molar ratio with respect to the products. A ketimine accumulates during the linear phase of product formation. This species is reactive since it is converted back to pyridoxal 5'-phosphate when the substrate is consumed. Superoxide anion and hydrogen peroxide are both generated during the catalytic cycles.
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?
D-tryptophan methyl ester + 1/2 O2
methyl 3-(1H-indol-3yl)-2-oxopropanoate + NH3
show the reaction diagram
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production depending on the nature of the substrate, and ammonia with concomitant O2 consumption in a 1:2 molar ratio with respect to the products
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?
glycine + O2
oxoacetic acid + NH3
show the reaction diagram
L-alanine + O2
2-oxo-propanoic acid + NH3
show the reaction diagram
L-DOPA + O2
?
show the reaction diagram
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?
L-glutamate + O2
2-oxopentanedioic acid + NH3
show the reaction diagram
L-phenylalanine + O2
2-oxo-3-phenylpropanoic acid + NH3
show the reaction diagram
L-tryptophan + O2
3-(1H-indol-3-yl)-2-oxopropanoic acid + NH3
show the reaction diagram
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50% of the activity with 3,4-dihydroxy-L-phenylalanine
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?
L-tyrosine + O2
3-(4-hydroxyphenyl)-2-oxopropanoic acid + NH3
show the reaction diagram
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80% of the activity with 3,4-dihydroxy-L-phenylalanine
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?
additional information
?
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The novelty in DDC is the possibility of catalyzing a reaction involving dioxygen although the enzyme lacks of any cofactor or metal related to O2 chemistry. The external aldimine intermediate undergoes a decarboxylation or a deprotonation leading to a quinonoid species, that is protonated at C4 producing the ketimine intermediate. Although it cannot be ruled out that this intermediate could be attacked by dioxygen, it seems much more likely, regarding enzymes proceeding through a carbanion chemistry on DDC, that the more electron dense quinonoid intermediate, in equilibrium with the ketimine, is reactive toward O2. Aerobiosis shifts the quinonoid-ketimine equilibrium toward quinonoid, while anaerobiosis shifts the equilibrium toward ketimine. The reaction between dioxygen and the quinonoid give rise directly to a superoxide anion and semiquinone. Superoxide is deprotonated and its anionic form is thus able to couple with the semiquinone giving rise to a peroxide species that is further protonated, and thus forming a hydroperoxy-pyridoxal 5'-phosphate intermediate. This rearranges to produce aldehyde, ammonia and hydrogen peroxide.
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
aromatic amine + 1/2 O2
aromatic aldehyde + NH3 + H2O
show the reaction diagram
-
oxidative deamination, unusual oxygen-consuming reaction catalyzed by the enzyme toward aromatic amines (serotonin, dopamine, and alpha-methyldopamine) and D-tryptophan methyl ester
production in equivalent amounts depending on the nature of the substrate, and ammonia with concomitant O2 consumption in a 1:2 molar ratio with respect to the products. A ketimine accumulates during the linear phase of product formation. This species is reactive since it is converted back to pyridoxal 5'-phosphate when the substrate is consumed. Superoxide anion and hydrogen peroxide are both generated during the catalytic cycles.
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?
D-tryptophan methyl ester + 1/2 O2
methyl 3-(1H-indol-3yl)-2-oxopropanoate + NH3
show the reaction diagram
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production depending on the nature of the substrate, and ammonia with concomitant O2 consumption in a 1:2 molar ratio with respect to the products
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?
L-DOPA + O2
?
show the reaction diagram
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?
additional information
?
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The novelty in DDC is the possibility of catalyzing a reaction involving dioxygen although the enzyme lacks of any cofactor or metal related to O2 chemistry. The external aldimine intermediate undergoes a decarboxylation or a deprotonation leading to a quinonoid species, that is protonated at C4 producing the ketimine intermediate. Although it cannot be ruled out that this intermediate could be attacked by dioxygen, it seems much more likely, regarding enzymes proceeding through a carbanion chemistry on DDC, that the more electron dense quinonoid intermediate, in equilibrium with the ketimine, is reactive toward O2. Aerobiosis shifts the quinonoid-ketimine equilibrium toward quinonoid, while anaerobiosis shifts the equilibrium toward ketimine. The reaction between dioxygen and the quinonoid give rise directly to a superoxide anion and semiquinone. Superoxide is deprotonated and its anionic form is thus able to couple with the semiquinone giving rise to a peroxide species that is further protonated, and thus forming a hydroperoxy-pyridoxal 5'-phosphate intermediate. This rearranges to produce aldehyde, ammonia and hydrogen peroxide.
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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no cofactor required
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-oxoglutarate
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3-(3,4-dihydroxyphenyl)-2-hydrazino-2-methyl propionic acid
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carbiDOPA, addition of 10 microM inhibitor to reaction mixtures (Y332F mutant with L-dopa) in the presence or in the absence of catalase or superoxide dismutase, immediately stops the O2 consumption.
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
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Km value for 3,4-dihydroxy-L-phenylalanine is 11.84 microM
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.68
3,4-dihydroxy-L-phenylalanine
Rhodobacter sphaeroides
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pH 7.8, 30°C
4.5
L-Dopa
Sus scrofa
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Y332F DDC mutant, reaction in 50 mM Hepes, pH 7.5, at 25°C causes the production of ammonia and 3,4-dihydroxyphenylacetaldehyde along with the consumption of molecular oxygen in a 1:2 molar ratio
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
Rhodobacter sphaeroides
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IC50 value for NADH is 35 microM per l, IC50 value for 2-oxoglutarate is 25 microM per l
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
190000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pentamer
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and expression of wild-type 3,4-dihydroxyphenylalanine oxidative deaminase and Y332F and T246A mutants in SVS370 Escherichia coli cells.
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
T246A
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T246 act as an essential general base for the oxidative deamination reaction
Y332F
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wild-type enzyme and Y332F variant are able to perform the oxidation toward aromatic amines or aromatic L-amino acids, without the aid of any cofactor related to oxygen chemistry.