Literature summary for 1.4.3.3 extracted from

Takahashi, S.; Furukawara, M.; Omae, K.; Tadokoro, N.; Saito, Y.; Abe, K.; Kera, Y.
A highly stable d-amino acid oxidase of the thermophilic bacterium Rubrobacter xylanophilus (2014), Appl. Environ. Microbiol., 80, 7219-7229 .

Search for more literature for 1.4.3.3

Activating Compound

Activating Compound	Comment	Organism	Structure
2,3-Butanedione	115% activity at 10 mM	Rubrobacter xylanophilus
D-malate	107% activity at 10 mM	Rubrobacter xylanophilus
iodoacetate	112% activity at 10 mM	Rubrobacter xylanophilus

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) or BL21 Star(DE3) cells	Rubrobacter xylanophilus

Inhibitors

Inhibitors	Comment	Organism	Structure
anthranilate	83.3% residual activity at 10 mM	Rubrobacter xylanophilus
benzoate	37% residual activity at 10 mM	Rubrobacter xylanophilus
crotonate	89.2% residual activity at 10 mM	Rubrobacter xylanophilus
D-Tyr	-	Rubrobacter xylanophilus
additional information	not inhibited by diethyl pyrocarbonate, malonate, and meso-tartrate	Rubrobacter xylanophilus
phenylmethylsulfonyl fluoride	31.8% residual activity at 10 mM	Rubrobacter xylanophilus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.049	-	D-Leu	at pH 8.0 and 60°C	Rubrobacter xylanophilus
0.052	-	D-Ile	at pH 8.0 and 60°C	Rubrobacter xylanophilus
0.094	-	D-Val	at pH 8.0 and 60°C	Rubrobacter xylanophilus
0.197	-	D-Tyr	at pH 8.0 and 60°C	Rubrobacter xylanophilus
8.16	-	D-Thr	at pH 8.0 and 60°C	Rubrobacter xylanophilus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
24100	-	gel filtration	Rubrobacter xylanophilus

Organism

Organism	UniProt	Comment	Textmining
Rubrobacter xylanophilus	A0A077JN67	-	-

Purification (Commentary)

Purification (Comment)	Organism
Talon metal affinity column chromatography, and Superdex 200 gel filtration	Rubrobacter xylanophilus

Storage Stability

Storage Stability	Organism
30°C, 1 week, no loss of activity	Rubrobacter xylanophilus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Ala + H2O + O2	1.6% activity compared to D-Val	Rubrobacter xylanophilus	pyruvate + NH3 + H2O2	-	?
D-Arg + H2O + O2	1.1% activity compared to D-Val	Rubrobacter xylanophilus	5-guanidino-2-oxopentanoate + NH3 + H2O2	-	?
D-Gln + H2O + O2	1.0% activity compared to D-Val	Rubrobacter xylanophilus	2-oxoglutaramate + NH3 + H2O2	-	?
D-Glu + H2O + O2	5.1% activity compared to D-Val	Rubrobacter xylanophilus	alpha-ketoglutarate + NH3 + H2O2	-	?
D-His + H2O + O2	3.6% activity compared to D-Val	Rubrobacter xylanophilus	3-(1H-imidazol-4-yl)-2-oxopropanoic acid + NH3 + H2O2	-	?
D-Ile + H2O + O2	26.7% activity compared to D-Val	Rubrobacter xylanophilus	3-methyl-2-oxopentanoate + NH3 + H2O2	-	?
D-Leu + H2O + O2	27.2% activity compared to D-Val	Rubrobacter xylanophilus	4-methyl-2-oxopentanoate + NH3 + H2O2	-	?
D-Lys + H2O + O2	2.4% activity compared to D-Val	Rubrobacter xylanophilus	6-amino-2-oxohexanoic acid + NH3 + H2O2	-	?
D-Met + H2O + O2	2.0% activity compared to D-Val	Rubrobacter xylanophilus	4-methylthio-2-oxobutanoate + NH3 + H2O2	-	?
D-Phe + H2O + O2	2.1% activity compared to D-Val	Rubrobacter xylanophilus	phenylpyruvate + NH3 + H2O2	-	?
D-Ser + H2O + O2	1.3% activity compared to D-Val	Rubrobacter xylanophilus	2-oxo-3-hydroxypropionate + NH3 + H2O2	-	?
D-Thr + H2O + O2	11.5% activity compared to D-Val	Rubrobacter xylanophilus	2-oxo-3-hydroxybutyrate + NH3 + H2O2	-	?
D-Trp + H2O + O2	1.5% activity compared to D-Val	Rubrobacter xylanophilus	indol-3-pyruvate + NH3 + H2O2	-	?
D-Tyr + H2O + O2	17.6% activity compared to D-Val	Rubrobacter xylanophilus	4-hydroxyphenylpyruvate + NH3 + H2O2	-	?
D-Val + H2O + O2	100% activity	Rubrobacter xylanophilus	2-oxoisovalerate + NH3 + H2O2	-	?
Gly + H2O + O2	1.1% activity compared to D-Val	Rubrobacter xylanophilus	acetate + NH3 + H2O2	-	?
additional information	no activity with D-Pro, D-Cys, D-Asp, L-Val, L-Leu, and L-Ile	Rubrobacter xylanophilus	?	-	?

Subunits

Subunits	Comment	Organism
monomer	1 * 35000, SDS-PAGE	Rubrobacter xylanophilus
monomer	1 * 35100, calculated from amino acid sequence	Rubrobacter xylanophilus

Synonyms

Synonyms	Comment	Organism
DAO	-	Rubrobacter xylanophilus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
65	-	-	Rubrobacter xylanophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
30	64	50% of the initial enzymatic activity is lost after 1h at 64°C. The half-lives of the enzyme are 462 h at 30°C and 114 h at 50°C	Rubrobacter xylanophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
7.6	-	D-Thr	at pH 8.0 and 60°C	Rubrobacter xylanophilus
23	-	D-Ile	at pH 8.0 and 60°C	Rubrobacter xylanophilus
24	-	D-Leu	at pH 8.0 and 60°C	Rubrobacter xylanophilus
31.4	-	D-Val	at pH 8.0 and 60°C	Rubrobacter xylanophilus
53	-	D-Tyr	at pH 8.0 and 60°C	Rubrobacter xylanophilus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	8	-	Rubrobacter xylanophilus

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
5	8	the enzyne is stable at a pH 5.0 to 8.0 for 1 h at 55°C	Rubrobacter xylanophilus

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Rubrobacter xylanophilus

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.265	-	D-Tyr	at pH 8.0 and 60°C	Rubrobacter xylanophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.931	-	D-Thr	at pH 8.0 and 60°C	Rubrobacter xylanophilus
269	-	D-Tyr	at pH 8.0 and 60°C	Rubrobacter xylanophilus
334	-	D-Val	at pH 8.0 and 60°C	Rubrobacter xylanophilus
442	-	D-Ile	at pH 8.0 and 60°C	Rubrobacter xylanophilus
490	-	D-Leu	at pH 8.0 and 60°C	Rubrobacter xylanophilus

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Release 2023.1 (January 2023)