P80324 · OXDA_RHOTO
- ProteinD-amino-acid oxidase
- GeneDAO1
- StatusUniProtKB reviewed (Swiss-Prot)
- Amino acids368 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Catalyzes the oxidative deamination of D-amino acids with broad substrate specificity (PubMed:12021281, PubMed:12208501, PubMed:15058991, PubMed:15310841, PubMed:17145127, PubMed:18343336, PubMed:19694805, PubMed:21053050, PubMed:26132227, PubMed:34174417, PubMed:8100225, PubMed:9383984, Ref.14). Enables the organism to utilize D-amino acids as a source of nutrients (By similarity).
Catalytic activity
- a D-alpha-amino acid + H2O + O2 = a 2-oxocarboxylate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-leucine + H2O + O2 = 4-methyl-2-oxopentanoate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-alanine + H2O + O2 = H2O2 + NH4+ + pyruvateThis reaction proceeds in the forward direction.
- D-glutamate + H2O + O2 = 2-oxoglutarate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-histidine + H2O + O2 = 3-(imidazol-5-yl)pyruvate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-proline + O2 = 1-pyrroline-2-carboxylate + H2O2This reaction proceeds in the forward direction.
- D-methionine + H2O + O2 = 4-methylsulfanyl-2-oxobutanoate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-arginine + H2O + O2 = 5-guanidino-2-oxopentanoate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-valine + H2O + O2 = 3-methyl-2-oxobutanoate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-threonine + H2O + O2 = (S)-3-hydroxy-2-oxobutanoate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-serine + H2O + O2 = 3-hydroxypyruvate + H2O2 + NH4+This reaction proceeds in the forward direction.
- D-tryptophan + H2O + O2 = H2O2 + indole-3-pyruvate + NH4+This reaction proceeds in the forward direction.
Cofactor
Activity regulation
Inhibited by high concentrations of hydrogen peroxide.
Biotechnology
Capable of converting cephalosporin C (CPC) into 7-beta-(5-carboxy-5-oxopentanamido)-cephalosporinic acid, an initial material of different beta-lactam antibiotics (PubMed:17145127, PubMed:7654197, PubMed:9383984).
Has been used as a biosensor to detect and quantify D-amino acids; when the technology was implanted in the cortex of rats, the increase in the concentration of D-serine following an intraperitoneal injection could be sensed (PubMed:18229946).
Has been used to resolve racemic mixtures of D,L-amino acids to yield enantiomerically pure isomers for pharmacological and peptide research uses (Ref.14). Has potential uses in anticancer treatment strategies; a recombinant enzyme with improved activity at low oxygen and substrate concentrations is targeted to tumor cells where it oxidizes D-amino acid prodrugs, yielding the cytotoxic metabolite hydrogen peroxide (PubMed:19694805, PubMed:34174417, PubMed:9472778).
Has been used as a biosensor to detect and quantify D-amino acids; when the technology was implanted in the cortex of rats, the increase in the concentration of D-serine following an intraperitoneal injection could be sensed (PubMed:18229946).
Has been used to resolve racemic mixtures of D,L-amino acids to yield enantiomerically pure isomers for pharmacological and peptide research uses (Ref.14). Has potential uses in anticancer treatment strategies; a recombinant enzyme with improved activity at low oxygen and substrate concentrations is targeted to tumor cells where it oxidizes D-amino acid prodrugs, yielding the cytotoxic metabolite hydrogen peroxide (PubMed:19694805, PubMed:34174417, PubMed:9472778).
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
0.9 mM | D-alanine using free enzyme | 8 | 25 | |||
1.6 mM | D-alanine using immobilized enzyme | 8 | 25 | |||
0.8 mM | D-alanine | 8.5 | 25 | |||
0.9 mM | D-alanine | 8.5 | 25 | |||
2.6 mM | D-alanine | 8.5 | 15 | |||
1 mM | D-alanine | 8.5 | 25 | |||
21.5 mM | D-proline | 8.5 | 25 | |||
14.4 mM | D-asparagine | 8.5 | 25 | |||
4.6 mM | D-glutamine | 8.5 | 25 | |||
0.2 mM | D-methionine | 8.5 | 25 | |||
18 mM | D-aspartate | 8.5 | 25 | |||
33.1 mM | D-aspartate | 8.5 | 25 | |||
77.3 mM | D-glutamate | 8.5 | 25 | |||
2.8 mM | D-arginine | 8.5 | 25 | |||
18 mM | D-arginine | 8.5 | 25 | |||
5 mM | cephalosporin C (CPC) | 8.5 | 25 | |||
4 mM | cephalosporin C (CPC) | 8.5 | 25 | |||
0.04 mM | 3-(2-Naphthyl)-D-alanine | 8.5 | 25 | |||
0.04 mM | 3-(1-Naphthyl)-D-alanine | 8.5 | 25 | |||
0.01 mM | d-2-Naphthylglycine | 8.5 | 25 | |||
0.33 mM | d-1-Naphthylglycine | 8.5 | 25 | |||
1.9 mM | oxygen | 8.5 | 15 |
kcat is 52.4 sec-1 with D-alanine as substrate and with free enzyme (at 25 degrees Celsius and at pH 8) (PubMed:18343336).
kcat is 23.3 sec-1 with D-alanine as substrate and with immobilized enzyme (at 25 degrees Celsius and at pH 8) (PubMed:18343336).
kcat is 83 sec-1 with D-alanine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 81 sec-1 with D-alanine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 330 sec-1 with D-alanine as substrate and with free enzyme (at 15 degrees Celsius and at pH 8.5) (PubMed:19694805).
kcat is 81.3 sec-1 with D-alanine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:15058991).
kcat is 77.3 sec-1 with D-proline as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 21.6 sec-1 with D-asparagine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 18.9 sec-1 with D-glutamine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 76 sec-1 with D-methionine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 20 sec-1 with D-arginine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 0.5 sec-1 with D-aspartate as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 0.5 sec-1 with D-aspartate as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 1 sec-1 with D-glutamate as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 72 sec-1 with cephalosporin C (CPC) as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 73.3 sec-1 with cephalosporin C (CPC) as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:15058991).
kcat is 23.3 sec-1 with D-alanine as substrate and with immobilized enzyme (at 25 degrees Celsius and at pH 8) (PubMed:18343336).
kcat is 83 sec-1 with D-alanine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 81 sec-1 with D-alanine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 330 sec-1 with D-alanine as substrate and with free enzyme (at 15 degrees Celsius and at pH 8.5) (PubMed:19694805).
kcat is 81.3 sec-1 with D-alanine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:15058991).
kcat is 77.3 sec-1 with D-proline as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 21.6 sec-1 with D-asparagine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 18.9 sec-1 with D-glutamine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 76 sec-1 with D-methionine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 20 sec-1 with D-arginine as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 0.5 sec-1 with D-aspartate as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 0.5 sec-1 with D-aspartate as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 1 sec-1 with D-glutamate as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:12021281).
kcat is 72 sec-1 with cephalosporin C (CPC) as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:17145127).
kcat is 73.3 sec-1 with cephalosporin C (CPC) as substrate (at 25 degrees Celsius and at pH 8.5) (PubMed:15058991).
pH Dependence
Optimum pH is 8-10.
Temperature Dependence
Optimum temperature is 50 degrees Celsius.
Features
Showing features for binding site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 12 | FAD (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 15 | FAD (UniProtKB | ChEBI) | ||||
Sequence: I | ||||||
Binding site | 35 | FAD (UniProtKB | ChEBI) | ||||
Sequence: R | ||||||
Binding site | 36 | FAD (UniProtKB | ChEBI) | ||||
Sequence: D | ||||||
Binding site | 47 | FAD (UniProtKB | ChEBI) | ||||
Sequence: A | ||||||
Binding site | 48 | FAD (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 52 | FAD (UniProtKB | ChEBI) | ||||
Sequence: G | ||||||
Binding site | 54 | FAD (UniProtKB | ChEBI) | ||||
Sequence: N | ||||||
Binding site | 58 | anthranilate 2 (UniProtKB | ChEBI) | ||||
Sequence: F | ||||||
Binding site | 162 | FAD (UniProtKB | ChEBI) | ||||
Sequence: V | ||||||
Binding site | 223 | (R)-lactate (UniProtKB | ChEBI) | ||||
Sequence: Y | ||||||
Binding site | 223 | anthranilate 1 (UniProtKB | ChEBI) | ||||
Sequence: Y | ||||||
Binding site | 238 | (R)-lactate (UniProtKB | ChEBI) | ||||
Sequence: Y | ||||||
Binding site | 238 | anthranilate 2 (UniProtKB | ChEBI) | ||||
Sequence: Y | ||||||
Binding site | 285 | (R)-lactate (UniProtKB | ChEBI) | ||||
Sequence: R | ||||||
Binding site | 285 | anthranilate 1 (UniProtKB | ChEBI) | ||||
Sequence: R | ||||||
Binding site | 285 | FAD (UniProtKB | ChEBI) | ||||
Sequence: R | ||||||
Binding site | 334 | FAD (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 335 | (R)-lactate (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 335 | anthranilate 1 (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 335 | FAD (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 337 | FAD (UniProtKB | ChEBI) | ||||
Sequence: G | ||||||
Binding site | 338 | FAD (UniProtKB | ChEBI) | ||||
Sequence: Y | ||||||
Binding site | 339 | FAD (UniProtKB | ChEBI) | ||||
Sequence: Q |
GO annotations
Aspect | Term | |
---|---|---|
Cellular Component | peroxisomal matrix | |
Cellular Component | peroxisome | |
Molecular Function | D-amino-acid oxidase activity | |
Molecular Function | D-glutamate oxidase activity | |
Molecular Function | FAD binding | |
Biological Process | D-amino acid catabolic process | |
Biological Process | D-amino acid metabolic process | |
Biological Process | nitrogen utilization |
Keywords
- Molecular function
- Ligand
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameD-amino-acid oxidase
- EC number
- Short namesDAAO ; DAMOX; DAO
Gene names
Organism names
- Strains
- Taxonomic lineageEukaryota > Fungi > Dikarya > Basidiomycota > Pucciniomycotina > Microbotryomycetes > Sporidiobolales > Sporidiobolaceae > Rhodotorula
Accessions
- Primary accessionP80324
Subcellular Location
Phenotypes & Variants
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Mutagenesis | 19 | Increases activity at low oxygen and D-alanine concentrations; when associated with P-120; R-144; M-321 and V-345. | ||||
Sequence: S → G | ||||||
Mutagenesis | 60 | Increases activity on D-arginine; when associated with R-144 and E-152. | ||||
Sequence: T → A | ||||||
Mutagenesis | 118 | Increases activity on D-arginine. | ||||
Sequence: L → H | ||||||
Mutagenesis | 120 | Increases activity at low oxygen and D-alanine concentrations; when associated with G-19; R-144; M-321 and V-345. | ||||
Sequence: S → P | ||||||
Mutagenesis | 144 | Increases activity on D-aspartate. Increases activity at low oxygen and D-alanine concentrations. Increases activity on D-alanine; when associated with D-199; C-223 and R-329 or when associated with G-19; P-120; M-321 and V-345. Increases activity on D-arginine; when associated with A-60 and E-152. | ||||
Sequence: Q → R | ||||||
Mutagenesis | 152 | Increases activity on D-arginine; when associated with A-60 and R-144. | ||||
Sequence: K → E | ||||||
Mutagenesis | 199 | Increases activity on D-alanine; when associated with R-144; D-199; C-223 and R-329. | ||||
Sequence: G → D | ||||||
Mutagenesis | 213 | Alters substrate specificity; capable of oxidizing D-amino acids with an acidic side-chain. Increases activity on D-aspartate. | ||||
Sequence: M → E | ||||||
Mutagenesis | 213 | Alters substrate specificity. | ||||
Sequence: M → G or R | ||||||
Mutagenesis | 213 | Capable of oxidizing D-amino acids with an acidic side-chain and increases activity on D-aspartate. | ||||
Sequence: M → R | ||||||
Mutagenesis | 223 | Increases activity on D-alanine; when associated with R-144; D-199 and R-329. | ||||
Sequence: Y → C | ||||||
Mutagenesis | 223 | Decreases activity. | ||||
Sequence: Y → F or S | ||||||
Mutagenesis | 238 | Decreases activity. | ||||
Sequence: Y → F or S | ||||||
Mutagenesis | 238 | Alters substrate specificity; capable of oxidizing D-amino acids with an acidic side-chain. Increases activity on D-aspartate. | ||||
Sequence: Y → R | ||||||
Mutagenesis | 242 | Increases activity on D-arginine; when associated with R-253 and V-304. | ||||
Sequence: D → V | ||||||
Mutagenesis | 253 | Increases activity on D-arginine; when associated with V-242 and V-304. | ||||
Sequence: Q → R | ||||||
Mutagenesis | 304 | Increases activity on D-arginine; when associated with V-242 and R-253. | ||||
Sequence: D → V | ||||||
Mutagenesis | 308-321 | Loss of dimerization. | ||||
Sequence: Missing | ||||||
Mutagenesis | 321 | Increases activity at low oxygen and D-alanine concentrations; when associated with G-19; P-120; R-144 and V-345. | ||||
Sequence: K → M | ||||||
Mutagenesis | 329 | Increases activity on D-alanine; when associated with R-144; D-199 and C-223. | ||||
Sequence: H → R | ||||||
Mutagenesis | 335 | Decreases activity. | ||||
Sequence: S → G or R | ||||||
Mutagenesis | 339 | Decreases activity. | ||||
Sequence: Q → E or N | ||||||
Mutagenesis | 345 | Increases activity at low oxygen and D-alanine concentrations; when associated with G-19; P-120; R-144 and M-321. | ||||
Sequence: A → V |
PTM/Processing
Features
Showing features for signal, chain, glycosylation.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Signal | 1-24 | |||||
Sequence: MHSQKRVVVLGSGVIGLSSALILA | ||||||
Chain | PRO_0000162767 | 25-368 | D-amino-acid oxidase | |||
Sequence: RKGYSVHILARDLPEDVSSQTFASPWAGANWTPFMTLTDGPRQAKWEESTFKKWVELVPTGHAMWLKGTRRFAQNEDGLLGHWYKDITPNYRPLPSSECPPGAIGVTYDTLSVHAPKYCQYLARELQKLGATFERRTVTSLEQAFDGADLVVNATGLGAKSIAGIDDQAAEPIRGQTVLVKSPCKRCTMDSSDPASPAYIIPRPGGEVICGGTYGVGDWDLSVNPETVQRILKHCLRLDPTISSDGTIEGIEVLRHNVGLRPARRGGPRVEAERIVLPLDRTKSPLSLGRGSARAAKEKEVTLVHAYGFSSAGYQQSWGAAEDVAQLVDEAFQRYHGAARESKL | ||||||
Glycosylation | 177 | N-linked (GlcNAc...) asparagine | ||||
Sequence: N |
Keywords
- PTM
Expression
Induction
Induced by D-alanine.
Interaction
Subunit
Homodimer.
Structure
Family & Domains
Features
Showing features for region, motif.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Region | 308-321 | Required for dimerization | ||||
Sequence: SPLSLGRGSARAAK | ||||||
Motif | 366-368 | Microbody targeting signal | ||||
Sequence: SKL |
Sequence similarities
Belongs to the DAMOX/DASOX family.
Keywords
- Domain
Family and domain databases
Sequence
- Sequence statusComplete
- Sequence processingThe displayed sequence is further processed into a mature form.
- Length368
- Mass (Da)40,076
- Last updated1995-11-01 v1
- Checksum99940118BCFA886E
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
U60066 EMBL· GenBank· DDBJ | AAB51107.1 EMBL· GenBank· DDBJ | mRNA | ||
Z71657 EMBL· GenBank· DDBJ | CAA96323.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
AF003339 EMBL· GenBank· DDBJ | AAB93974.1 EMBL· GenBank· DDBJ | mRNA | ||
AF003340 EMBL· GenBank· DDBJ | AAB93975.1 EMBL· GenBank· DDBJ | Genomic DNA |