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Literature summary for 1.4.3.3 extracted from

  • Kuan, I.; Liao, R.; Hsieh, H.; Chen, K.; Yu, C.
    Properties of Rhodotorula gracilis D-amino acid oxidase immobilized on magnetic beads through His-tag (2008), J. Biosci. Bioeng., 105, 110-115.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Rhodotorula toruloides

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.9
-
D-alanine free enzyme, in 100 mM potassium phosphate buffer at pH 8.0 and 25°C Rhodotorula toruloides
1.6
-
D-alanine immobilized enzyme, in 100 mM potassium phosphate buffer at pH 8.0 and 25°C Rhodotorula toruloides

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
2 * 40000, SDS-PAGE Rhodotorula toruloides
80000
-
SDS-PAGE Rhodotorula toruloides

Organism

Organism UniProt Comment Textmining
Rhodotorula toruloides
-
-
-

Oxidation Stability

Oxidation Stability Organism
the immobilized enzyme (onto Ni2+-chelated NTA magnetic beads) does not show any loss of activity in the presence of 10 mM H2O2 after 2 h incubation which is observed in case of free enzyme, the residual activity of the immoblized enzyme after 9 h is 72% compared with 22% of the free enzyme Rhodotorula toruloides

Purification (Commentary)

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

Source Tissue

Source Tissue Comment Organism Textmining

Storage Stability

Storage Stability Organism
22°C, free enzyme, 10 days, 20% residual activity Rhodotorula toruloides
22°C, immobilized enzyme, 10 days, 74% residual activity Rhodotorula toruloides

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cephalosporin C + H2O + O2
-
Rhodotorula toruloides 7-(5-oxoadipoamido)cephalosporanic acid + NH3 + H2O2
-
?
D-alanine + H2O + O2
-
Rhodotorula toruloides pyruvate + NH3 + H2O2
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 40000, SDS-PAGE Rhodotorula toruloides

Synonyms

Synonyms Comment Organism
D-amino acid oxidase
-
Rhodotorula toruloides
DAO
-
Rhodotorula toruloides

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
free and immobilized enzymes Rhodotorula toruloides

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
the native enzyme is inacivated after 1 h at 50°C whereas the immobilized enzyme (onto Ni2+-chelated NTA magnetic beads) retains 56% of the initial activity Rhodotorula toruloides

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
23.3
-
D-alanine immobilized enzyme, in 100 mM potassium phosphate buffer at pH 8.0 and 25°C Rhodotorula toruloides
52.4
-
D-alanine free enzyme, in 100 mM potassium phosphate buffer at pH 8.0 and 25°C Rhodotorula toruloides

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
free enzyme Rhodotorula toruloides
9
-
immobilized enzyme Rhodotorula toruloides

pH Stability

pH Stability pH Stability Maximum Comment Organism
8 9 at pH 8 immobilized enzyme (onto Ni2+-chelated NTA magnetic beads) activity decreases to 44% while the free enzyme showed optimal activity, the activities of both enzyme forms decay similarily at pH values above 9, the free enzyme activity is less affected for pH values below 9 than the immobilized enzyme Rhodotorula toruloides

Cofactor

Cofactor Comment Organism Structure
FAD
-
Rhodotorula toruloides