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Literature summary extracted from

  • Piubelli, L.; Caldinelli, L.; Molla, G.; Pilone, M.S.; Pollegioni, L.
    Conversion of the dimeric D-amino acid oxidase from Rhodotorula gracilis to a monomeric form. A rational mutagenesis approach (2002), FEBS Lett., 526, 43-48.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.3.3 expression in Escherichia coli Rhodotorula toruloides

Protein Variants

EC Number Protein Variants Comment Organism
1.4.3.3 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 Rhodotorula toruloides

General Stability

EC Number General Stability Organism
1.4.3.3 incubation of the mutant enzyme DELTASer308-Lys321 with 10% trypsin in presence of exogenous FAD, 0.2 mM, shows that the rate of activity loss is lower than that determined in absence of exogenous FAD Rhodotorula toruloides

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.3.3 0.4
-
 D-Trp pH 7.5, 25°C, mutant enzyme DELTASer308-Lys321 Rhodotorula toruloides
1.4.3.3 1.5
-
 D-Ala pH 7.5, 25°C, mutant enzyme DELTASer308-Lys321 Rhodotorula toruloides

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.3.3 37200
-
 gel filtration Rhodotorula toruloides
1.4.3.3 40000
-
 1 * 40000, mutant enzyme DELTASer308-Lys321, SDS-PAGE Rhodotorula toruloides

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.3 Rhodotorula toruloides P80324
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.3
-
 Rhodotorula toruloides

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.3 D-alanine + H2O + O2
-
 Rhodotorula toruloides pyruvate + NH3 + H2O2
-
 ?
1.4.3.3 D-tryptophan + H2O + O2
-
 Rhodotorula toruloides indol-3-pyruvic acid + NH3 + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.3.3 monomer 1 * 40000, mutant enzyme DELTASer308-Lys321, SDS-PAGE Rhodotorula toruloides

Synonyms

EC Number Synonyms Comment Organism
1.4.3.3 RgDAAO
-
 Rhodotorula toruloides

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.3.3 40
-
 30 min, wild-type holoenzym retains 60% of the initial activity, wild-type apoprotein retains 20% of the initial activity, mutant holoenzyme DELTASer308-Lys321 retains less than 5% of the initial activity Rhodotorula toruloides

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.3.3 8.5
-
 wild-type enzyme and mutant enzyme DELTASer308-Lys321 Rhodotorula toruloides

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.4.3.3 6 9 wild-type enzyme and mutant enzyme DELTASer308-Lys321 are stable betweeen Rhodotorula toruloides

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.3 FAD mutant enzyme DELTASer308-Lys321 retains the binding of the FAD coenzyme, binding of mutant apoenzyme is weaker than that of wild-type apoenzyme to FAD Rhodotorula toruloides

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.4.3.3 Rhodotorula toruloides and pI 7.6, isoelectrofocusing, wild-type enzyme 4.7 4.6
1.4.3.3 Rhodotorula toruloides and pI 6.8, isoelectrofocusing, mutant enzyme DELTASer308-Lys321
-
 6.9
1.4.3.3 Rhodotorula toruloides and pI 8.8, isoelectrofocusing, wild-type enzyme
-
 6.9
1.4.3.3 Rhodotorula toruloides and pI 8.2, isoelectrofocusing, mutant enzyme DELTASer308-Lys321
-
 7.1