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Literature summary for 1.4.3.3 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)

Cloned (Comment) Organism
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
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]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Rhodotorula toruloides
-
-
-
Trigonopsis variabilis
-
-
-

Oxidation Stability

Oxidation Stability Organism
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
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)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

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

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

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

pH Optimum

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

pH Stability

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

Cofactor

Cofactor Comment Organism Structure
FAD
-
Rhodotorula toruloides
FAD
-
Trigonopsis variabilis