Literature summary for 1.4.3.16 extracted from

Leese, C.; Fotheringham, I.; Escalettes, F.; Speight, R.; Grogan, G.
Cloning, expression, characterisation and mutational analysis of L-aspartate oxidase from Pseudomonas putida (2013), J. Mol. Catal. B, 85-86, 17-22.

No PubMed abstract available

Search for more literature for 1.4.3.16

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21-Gold (DE3) cells	Pseudomonas putida

Protein Variants

Protein Variants	Comment	Organism
H244A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Pseudomonas putida
H351A	inactive	Pseudomonas putida
Q242A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Pseudomonas putida
R290A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Pseudomonas putida
R386A	inactive	Pseudomonas putida
S389A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Pseudomonas putida
T259A	inactive	Pseudomonas putida

Inhibitors

Inhibitors	Comment	Organism	Structure
L-asparagine	-	Pseudomonas putida

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
2.26	-	L-aspartate	wild type enzyme, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
14.25	-	L-aspartate	mutant enzyme S389A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
26.11	-	L-aspartate	mutant enzyme R290A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
63.27	-	L-aspartate	mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
75.44	-	L-aspartate	mutant enzyme H244A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
59900	-	x * 59900, calculated from amino acid sequence	Pseudomonas putida
60000	-	x * 60000, SDS-PAGE	Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-asparagine + H2O + O2	Pseudomonas putida	low activity	4-amino-2,4-dioxobutanoate + NH3 + H2O2	-	?
L-aspartate + H2O + O2	Pseudomonas putida	best substrate	oxaloacetate + NH3 + H2O2	-	?
L-glutamate + H2O + O2	Pseudomonas putida	low activity	2-oxopentanedioate + NH3 + H2O2	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
HiTrap nickel chelating column chromatography and Superdex 75 gel filtration	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-asparagine + H2O + O2	low activity	Pseudomonas putida	4-amino-2,4-dioxobutanoate + NH3 + H2O2	-	?
L-aspartate + H2O + O2	best substrate	Pseudomonas putida	oxaloacetate + NH3 + H2O2	-	?
L-glutamate + H2O + O2	low activity	Pseudomonas putida	2-oxopentanedioate + NH3 + H2O2	-	?

Subunits

Subunits	Comment	Organism
?	x * 60000, SDS-PAGE	Pseudomonas putida
?	x * 59900, calculated from amino acid sequence	Pseudomonas putida

Synonyms

Synonyms	Comment	Organism
LASPO	-	Pseudomonas putida

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
40	50	the enzyme is stable for 60 min at up to 40°C, but rapid losses in activity are observed at 50°C after 5 min	Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.23	-	L-aspartate	mutant enzyme R290A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
2.28	-	L-aspartate	mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
7.83	-	L-aspartate	mutant enzyme S389A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
10.6	-	L-aspartate	wild type enzyme, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
23.69	-	L-aspartate	mutant enzyme H244A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	-	Pseudomonas putida

pH Range

pH Minimum	pH Maximum	Comment	Organism
6	8	about 40% activity at pH 6.0, about 50% activity at pH 8.0	Pseudomonas putida

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Pseudomonas putida

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.00884	-	L-aspartate	mutant enzyme R290A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
0.036	-	L-aspartate	mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
0.314	-	L-aspartate	mutant enzyme H244A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
0.549	-	L-aspartate	mutant enzyme S389A, at pH 7.5, temperature not specified in the publication	Pseudomonas putida
4.69	-	L-aspartate	wild type enzyme, at pH 7.5, temperature not specified in the publication	Pseudomonas putida

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)