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Literature summary for 3.5.3.1 extracted from

  • Garcia, D.; Uribe, E.; Salgado, M.; Martinez, M.P.; Carvajal, N.
    Mutagenic and kinetic support for an allosteric site in arginase from the extreme thermophile Bacillus caldovelox, which allows activation by arginine (2015), Biochimie, 108, 8-12 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
L-arginine the enzyme is kinetically cooperative and the commonly detected hyperbolic behavior results from binding of L-arginine as a typical allosteric activator [Bacillus] caldovelox

Protein Variants

Protein Variants Comment Organism
D199A in contrast with the Michaelis-Menten kinetics exhibited by the wild-type enzyme, the mutant enzyme exhibits positive cooperativity with respect to L-arginine. Mutation has no effect on the hexameric structure of the enzyme [Bacillus] caldovelox
D199N in contrast with the Michaelis-Menten kinetics exhibited by the wild-type enzyme, the mutant enzyme exhibits positive cooperativity with respect to L-arginine. Mutation has no effect on the hexameric structure of the enzyme [Bacillus] caldovelox
E256A in contrast with the Michaelis-Menten kinetics exhibited by the wild-type enzyme, the mutant enzyme exhibits positive cooperativity with respect to L-arginine. Mutation has no effect on the hexameric structure of the enzyme [Bacillus] caldovelox
E256A/E278A sigmoidal kinetics [Bacillus] caldovelox
E256Q in contrast with the Michaelis-Menten kinetics exhibited by the wild-type enzyme, the mutant enzyme exhibits positive cooperativity with respect to L-arginine. Mutation has no effect on the hexameric structure of the enzyme [Bacillus] caldovelox
E278A the mutation compromises interactions at the trimer/trimer interface and yields trimeric species (MW 100000 Da) exhibiting hyperbolic kinetics that changed to sigmoidal by the additional E256A mutation [Bacillus] caldovelox

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4
-
L-arginine pH 9.5, 55°C, wild-type enzyme [Bacillus] caldovelox
5
-
L-arginine pH 9.5, 55°C, mutant enzyme E278A [Bacillus] caldovelox
6.3
-
L-arginine pH 9.5, 55°C, mutant enzyme E256A [Bacillus] caldovelox
6.5
-
L-arginine pH 9.5, 55°C, mutant enzyme D199N [Bacillus] caldovelox
7.2
-
L-arginine pH 9.5, 55°C, mutant enzyme E256A/E278A [Bacillus] caldovelox
9.1
-
L-arginine pH 9.5, 55°C, mutant enzyme E256Q [Bacillus] caldovelox
9.8
-
L-arginine pH 9.5, 55°C, mutant enzyme D199A [Bacillus] caldovelox

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ all recombinant variants are active even in the absence of added Mn2+ but express about2.5-fold increased activity in the presence of 2 mM Mn2+. After extensive dialysis against 25 mM EDTA, they became totally dependent on added Mn2+ for catalytic activity. The manganese-reactivation of all fully inactivated species followed hyperbolic kinetics [Bacillus] caldovelox

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
195000
-
gel filtration [Bacillus] caldovelox

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arginine + H2O [Bacillus] caldovelox
-
L-ornithine + urea
-
?

Organism

Organism UniProt Comment Textmining
[Bacillus] caldovelox P53608
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arginine + H2O
-
[Bacillus] caldovelox L-ornithine + urea
-
?

Subunits

Subunits Comment Organism
hexamer 6 * 32000, SDS-PAGE [Bacillus] caldovelox

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
300
-
L-arginine pH 9.5, 55°C, mutant enzyme E256A [Bacillus] caldovelox
300
-
L-arginine pH 9.5, 55°C, mutant enzyme E256A/E278A [Bacillus] caldovelox
400
-
L-arginine pH 9.5, 55°C, mutant enzyme D199N [Bacillus] caldovelox
400
-
L-arginine pH 9.5, 55°C, mutant enzyme E256Q [Bacillus] caldovelox
500
-
L-arginine pH 9.5, 55°C, mutant enzyme D199A [Bacillus] caldovelox
700
-
L-arginine pH 9.5, 55°C, wild-type enzyme [Bacillus] caldovelox
750
-
L-arginine pH 9.5, 55°C, mutant enzyme E278A [Bacillus] caldovelox

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
44
-
L-arginine pH 9.5, 55°C, mutant enzyme E256Q [Bacillus] caldovelox
47.8
-
L-arginine pH 9.5, 55°C, mutant enzyme E256A [Bacillus] caldovelox
51.1
-
L-arginine pH 9.5, 55°C, mutant enzyme D199A [Bacillus] caldovelox
61.6
-
L-arginine pH 9.5, 55°C, mutant enzyme D199N [Bacillus] caldovelox
206.8
-
L-arginine pH 9.5, 55°C, wild-type enzyme [Bacillus] caldovelox