Literature summary for 1.4.3.3 extracted from

Rosini, E.; Pollegioni, L.; Ghisla, S.; Orru, R.; Molla, G.
Optimization of D-amino acid oxidase for low substrate concentrations - towards a cancer enzyme therapy (2009), FEBS J., 276, 4921-4932.

Search for more literature for 1.4.3.3

Application

Application	Comment	Organism
medicine	potential in vivo applicability of this evolved mutant DAAO, with increased activity at low O2 and D-Ala concentrations and a 10fold lower Km for O2, for tumor therapy	Rhodotorula toruloides
synthesis	the enzyme might be useful as a biocatalyst for industrial applications and for therapeutic treatments, mechanism of this DAAO variant and on its cytotoxicity towards various mammalian cancer cell lines, overview	Rhodotorula toruloides

Cloned(Commentary)

Cloned (Comment)	Organism
cDNA library screening	Rhodotorula toruloides

Protein Variants

Protein Variants	Comment	Organism
Q144R	site-directed mutagenesis, the mutant shows reduced activity comapared to the wild-type enzyme	Rhodotorula toruloides
S19G/S120P/Q144R/K321M/A345V	library screening and identification of a naturally occuring DAAO mutant with increased activity at low O2 and D-Ala concentrations and a 10fold lower Km for O2	Rhodotorula toruloides

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state kinetics, reductive and oxidative half-reactions, overview	Rhodotorula toruloides
1.4	-	O2	pH 8.5, 15°C, mutant Q144R	Rhodotorula toruloides
1.8	-	D-alanine	pH 8.5, 15°C, mutant Q144R	Rhodotorula toruloides
2.6	-	D-alanine	pH 8.5, 15°C, wild-type enzyme	Rhodotorula toruloides
4.7	-	D-alanine	pH 8.5, 15°C, mutant S19G/S120P/Q144R/K321M/A345V	Rhodotorula toruloides
5	-	O2	pH 8.5, 15°C, wild-type enzyme	Rhodotorula toruloides
6.1	-	O2	pH 8.5, 15°C, mutant S19G/S120P/Q144R/K321M/A345V	Rhodotorula toruloides

Organism

Organism	UniProt	Comment	Textmining
Rhodotorula toruloides	P80324	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
flavoprotein	the enzyme contains FAD as cofactor	Rhodotorula toruloides

Reaction

Reaction	Comment	Organism	Reaction ID
a D-amino acid + H2O + O2 = a 2-oxo carboxylate + NH3 + H2O2	catalytic mechanism, reductive half-reaction mechanism, wild-type and S19G/S120P/Q144R/K321M/A345V mutant enzymes, overview	Rhodotorula toruloides	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	-	Rhodotorula toruloides

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-alanine + H2O + O2	-	Rhodotorula toruloides	pyruvate + NH3 + H2O2	-	?

Synonyms

Synonyms	Comment	Organism
DAAO	-	Rhodotorula toruloides

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
15	-	assay at	Rhodotorula toruloides

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
140	-	D-alanine	pH 8.5, 15°C, mutant Q144R	Rhodotorula toruloides
330	-	D-alanine	pH 8.5, 15°C, wild-type enzyme	Rhodotorula toruloides
370	-	D-alanine	pH 8.5, 15°C, mutant S19G/S120P/Q144R/K321M/A345V	Rhodotorula toruloides

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	assay at	Rhodotorula toruloides

Cofactor

Cofactor	Comment	Organism	Structure
FAD	flavoenzyme	Rhodotorula toruloides

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Release 2023.1 (January 2023)