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Literature summary extracted from

  • Wang, S.J.; Yu, C.Y.; Lee, C.K.; Chern, M.K.; Kuan, I.C.
    Subunit fusion of two yeast D-amino acid oxidases enhances their thermostability and resistance to H2O2 (2008), Biotechnol. Lett., 30, 1415-1422.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.3.3 two cDNA genes were connected by a hexanucleotide linker and heterologously expressed in Escherichia coli to produce the corresponding double DAO with two subunits fused into a single polypeptide Rhodotorula toruloides
1.4.3.3 two cDNA genes were connected by a hexanucleotide linker and heterologously expressed in Escherichia coli to produce the corresponding double DAO with two subunits fused into a single polypeptide Trigonopsis variabilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.3.3 2
-
 D-alanine native enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 3
-
 D-alanine native enzyme, at 37°C Trigonopsis variabilis
1.4.3.3 6
-
 D-alanine double fusion enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 6
-
 cephalosporin C native enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 9
-
 cephalosporin C native enzyme, at 37°C Trigonopsis variabilis
1.4.3.3 13
-
 D-alanine double fusion enzyme, at 37°C Trigonopsis variabilis
1.4.3.3 13
-
 cephalosporin C double fusion enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 20
-
 cephalosporin C double fusion enzyme, at 37°C Trigonopsis variabilis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.3.3 20000
-
 2 * 20000, SDS-PAGE Rhodotorula toruloides
1.4.3.3 20000
-
 2 * 20000, SDS-PAGE Trigonopsis variabilis
1.4.3.3 40000
-
 SDS-PAGE Rhodotorula toruloides
1.4.3.3 40000
-
 SDS-PAGE Trigonopsis variabilis

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.3 Rhodotorula toruloides
-
-
-
1.4.3.3 Trigonopsis variabilis
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.4.3.3 double DAO exhibits a half-life of about 30 min in the presence of 10 mM H2O2 which is 1.5fold longer compared to the native DAO Trigonopsis variabilis
1.4.3.3 double DAO exhibits a half-life of about 60 min in the presence of 10 mM H2O2 which is 2fold longer compared to the native DAO Rhodotorula toruloides

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.3 His-Bind column chromatography Rhodotorula toruloides
1.4.3.3 His-Bind column chromatography Trigonopsis variabilis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.3.3 72
-
 native enzyme, with D-alanine as substrate, at 37°C Rhodotorula toruloides
1.4.3.3 77
-
 double fusion enzyme, with D-alanine as substrate, at 37°C Rhodotorula toruloides
1.4.3.3 95
-
 native enzyme, with D-alanine as substrate, at 37°C Trigonopsis variabilis
1.4.3.3 96
-
 double fusion enzyme, with D-alanine as substrate, at 37°C Trigonopsis variabilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.3 cephalosporin C + H2O + O2 14% activity compared to D-alanine Trigonopsis variabilis 7-(5-oxoadipoamido)cephalosporanic acid + NH3 + H2O2
-
 ?
1.4.3.3 cephalosporin C + H2O + O2 36% activity compared to D-alanine Rhodotorula toruloides 7-(5-oxoadipoamido)cephalosporanic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-alanine + H2O + O2 100% activity Rhodotorula toruloides pyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-alanine + H2O + O2 100% activity Trigonopsis variabilis pyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-arginine + H2O + O2 19% activity compared to D-alanine Rhodotorula toruloides 5-guanidino-2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-arginine + H2O + O2 42% activity compared to D-alanine Trigonopsis variabilis 5-guanidino-2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-asparagine + H2O + O2 38% activity compared to D-alanine Rhodotorula toruloides 2-oxosuccinamic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-asparagine + H2O + O2 67% activity compared to D-alanine Trigonopsis variabilis 2-oxosuccinamic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-aspartate + H2O + O2 2% activity compared to D-alanine Trigonopsis variabilis oxaloacetate + NH3 + H2O2
-
 ?
1.4.3.3 D-aspartate + H2O + O2 3% activity compared to D-alanine Rhodotorula toruloides oxaloacetate + NH3 + H2O2
-
 ?
1.4.3.3 D-cysteine + H2O + O2 18% activity compared to D-alanine Rhodotorula toruloides 2-oxo-3-thiopropionic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-cysteine + H2O + O2 9% activity compared to D-alanine Trigonopsis variabilis 2-oxo-3-thiopropionic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-glutamate + H2O + O2 3% activity compared to D-alanine Rhodotorula toruloides alpha-ketoglutarate + NH3 + H2O2
-
 ?
1.4.3.3 D-glutamate + H2O + O2 9% activity compared to D-alanine Trigonopsis variabilis alpha-ketoglutarate + NH3 + H2O2
-
 ?
1.4.3.3 D-glutamine + H2O + O2 58% activity compared to D-alanine Rhodotorula toruloides 2-oxoglutaramate + NH3 + H2O2
-
 ?
1.4.3.3 D-glutamine + H2O + O2 78% activity compared to D-alanine Trigonopsis variabilis 2-oxoglutaramate + NH3 + H2O2
-
 ?
1.4.3.3 D-histidine + H2O + O2 60% activity compared to D-alanine Rhodotorula toruloides 3-(1H-imidazol-4-yl)-2-oxopropanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-histidine + H2O + O2 69% activity compared to D-alanine Trigonopsis variabilis 3-(1H-imidazol-4-yl)-2-oxopropanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-isoleucine + H2O + O2 60% activity compared to D-alanine Rhodotorula toruloides 3-methyl-2-oxopentanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-isoleucine + H2O + O2 69% activity compared to D-alanine Trigonopsis variabilis 3-methyl-2-oxopentanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-leucine + H2O + O2 37% activity compared to D-alanine Trigonopsis variabilis 4-methyl-2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-leucine + H2O + O2 86% activity compared to D-alanine Rhodotorula toruloides 4-methyl-2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-lysine + H2O + O2 7% activity compared to D-alanine Trigonopsis variabilis 6-amino-2-oxohexanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-lysine + H2O + O2 8% activity compared to D-alanine Rhodotorula toruloides 6-amino-2-oxohexanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-methionine + H2O + O2 113% activity compared to D-alanine Rhodotorula toruloides 4-methylthio-2-oxobutanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-methionine + H2O + O2 77% activity compared to D-alanine Trigonopsis variabilis 4-methylthio-2-oxobutanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-phenylalanine + H2O + O2 44% activity compared to D-alanine Trigonopsis variabilis phenylpyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-phenylalanine + H2O + O2 94% activity compared to D-alanine Rhodotorula toruloides phenylpyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-proline + H2O + O2 21% activity compared to D-alanine Trigonopsis variabilis 2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-proline + H2O + O2 46% activity compared to D-alanine Rhodotorula toruloides 2-oxopentanoic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-serine + H2O + O2 23% activity compared to D-alanine Trigonopsis variabilis 2-oxo-3-hydroxypropionic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-serine + H2O + O2 49% activity compared to D-alanine Rhodotorula toruloides 2-oxo-3-hydroxypropionic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-threonine + H2O + O2 18% activity compared to D-alanine Rhodotorula toruloides 2-oxo-3-hydroxybutyric acid + NH3 + H2O2
-
 ?
1.4.3.3 D-threonine + H2O + O2 2% activity compared to D-alanine Trigonopsis variabilis 2-oxo-3-hydroxybutyric acid + NH3 + H2O2
-
 ?
1.4.3.3 D-tryptophan + H2O + O2 129% activity compared to D-alanine Trigonopsis variabilis indol-3-pyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-tryptophan + H2O + O2 92% activity compared to D-alanine Rhodotorula toruloides indol-3-pyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-tyrosine + H2O + O2 18% activity compared to D-alanine Rhodotorula toruloides 4-hydroxyphenylpyruvic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-tyrosine + H2O + O2 19% activity compared to D-alanine Trigonopsis variabilis 4-hydroxyphenylpyruvic acid + NH3 + H2O2
-
 ?
1.4.3.3 D-valine + H2O + O2 60% activity compared to D-alanine Rhodotorula toruloides alpha-ketoisovaleric acid + NH3 + H2O2
-
 ?
1.4.3.3 D-valine + H2O + O2 63% activity compared to D-alanine Trigonopsis variabilis alpha-ketoisovaleric acid + NH3 + H2O2
-
 ?
1.4.3.3 glycine + 2 H2O + O2 2% activity compared to D-alanine Trigonopsis variabilis 2 formic acid + NH3 + H2O2
-
 ?
1.4.3.3 glycine + 2 H2O + O2 3% activity compared to D-alanine Rhodotorula toruloides 2 formic acid + NH3 + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.3.3 homodimer 2 * 20000, SDS-PAGE Rhodotorula toruloides
1.4.3.3 homodimer 2 * 20000, SDS-PAGE Trigonopsis variabilis

Synonyms

EC Number Synonyms Comment Organism
1.4.3.3 D-amino acid oxidase
-
 Rhodotorula toruloides
1.4.3.3 D-amino acid oxidase
-
 Trigonopsis variabilis
1.4.3.3 DAAO
-
 Rhodotorula toruloides
1.4.3.3 DAAO
-
 Trigonopsis variabilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.3.3 50
-
 double fusion enzyme Rhodotorula toruloides
1.4.3.3 55
-
 native enzyme Rhodotorula toruloides
1.4.3.3 60
-
 double fusion enzyme Trigonopsis variabilis
1.4.3.3 65
-
 native enzyme Trigonopsis variabilis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.3.3 30 60 retains more than 80% of the maximal activity after incubation for 60 min at 30-60°C Rhodotorula toruloides
1.4.3.3 37 60 retains more than 80% of the maximal activity after incubation for 60 min at 37-60°C Trigonopsis variabilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.3.3 55
-
 cephalosporin C native enzyme, at 37°C Trigonopsis variabilis
1.4.3.3 60
-
 D-alanine native enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 64
-
 cephalosporin C double fusion enzyme, at 37°C Trigonopsis variabilis
1.4.3.3 70
-
 cephalosporin C native enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 74
-
 D-alanine native enzyme, at 37°C Trigonopsis variabilis
1.4.3.3 78
-
 D-alanine double fusion enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 80
-
 cephalosporin C double fusion enzyme, at 37°C Rhodotorula toruloides
1.4.3.3 88
-
 D-alanine double fusion enzyme, at 37°C Trigonopsis variabilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.3.3 8.5
-
 native and double fusion enzymes Rhodotorula toruloides
1.4.3.3 8.5
-
 native and double fusion enzymes Trigonopsis variabilis

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.4.3.3 5 10 retains more than 80% of the maximal activity after incubation for 60 min at pH 5-10 Rhodotorula toruloides
1.4.3.3 5 10 retains more than 80% of the maximal activity after incubation for 60 min at pH 5-10 Trigonopsis variabilis

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.3 FAD
-
 Rhodotorula toruloides
1.4.3.3 FAD
-
 Trigonopsis variabilis