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Literature summary for 3.5.3.6 extracted from

  • Lu, X.; Li, L.; Wu, R.; Feng, X.; Li, Z.; Yang, H.; Wang, C.; Guo, H.; Galkin, A.; Herzberg, O.; Mariano, P.S.; Martin, B.M.; Dunaway-Mariano, D.
    Kinetic analysis of Pseudomonas aeruginosa arginine deiminase mutants and alternate substrates provides insight into structural determinants of function (2006), Biochemistry, 45, 1162-1172.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pseudomonas aeruginosa
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
C406A inactive mutant enzyme Pseudomonas aeruginosa
C406A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
C406S inactive mutant enzyme Pseudomonas aeruginosa
C406S site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
D166A inactive mutant enzyme Pseudomonas aeruginosa
D166A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
D280A inactive mutant enzyme Pseudomonas aeruginosa
D280A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
E224A kcat/Km is 23684fold higher than the wild-type value Pseudomonas aeruginosa
E224A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
E224D kcat/Km is 50000fold higher than the wild-type value Pseudomonas aeruginosa
E224D site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
H278A inactive mutant enzyme Pseudomonas aeruginosa
H278A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
H278N inactive mutant enzyme Pseudomonas aeruginosa
H278N site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
H278V inactive mutant enzyme Pseudomonas aeruginosa
H278V site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
N160A kcat/Km is 5625fold higher than the wild-type value Pseudomonas aeruginosa
N160A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R185A kcat/Km is 7627fold higher than the wild-type value Pseudomonas aeruginosa
R185A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R185K site-directed mutagenesis, unstable mutant enzyme that precipitates Pseudomonas aeruginosa
R185L kcat/Km is 25000fold higher than the wild-type value Pseudomonas aeruginosa
R185L site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R243A kcat/Km is 150000fold higher than the wild-type value Pseudomonas aeruginosa
R243A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R243K kcat/Km is 150000fold higher than the wild-type value Pseudomonas aeruginosa
R243K site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R243L kcat/Km is 225000fold higher than the wild-type value Pseudomonas aeruginosa
R243L site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R401A kcat/Km is 56250fold higher than the wild-type value Pseudomonas aeruginosa
R401A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
R401L site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics Pseudomonas aeruginosa
0.14
-
L-arginine pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
0.14
-
L-Arg pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
1.1
-
L-Arg pH 5.6, 25°C, mutant enzyme E224A Pseudomonas aeruginosa
1.1
-
L-arginine pH 5.6, 25°C, mutant E224A Pseudomonas aeruginosa
2.6
-
L-Arg pH 5.6, 25°C, mutant enzyme R243A Pseudomonas aeruginosa
2.6
-
L-arginine pH 5.6, 25°C, mutant R243A Pseudomonas aeruginosa
2.7
-
L-Arg pH 5.6, 25°C, mutant enzyme N160A Pseudomonas aeruginosa
2.7
-
L-arginine pH 5.6, 25°C, mutant N160A Pseudomonas aeruginosa
4.2
-
L-Arg pH 5.6, 25°C, mutant enzyme E224D Pseudomonas aeruginosa
4.2
-
L-arginine pH 5.6, 25°C, mutant E224D Pseudomonas aeruginosa
4.8
-
L-Arg pH 5.6, 25°C, mutant enzyme R185L Pseudomonas aeruginosa
4.8
-
L-arginine pH 5.6, 25°C, mutant R185L Pseudomonas aeruginosa
6.9
-
L-Arg pH 5.6, 25°C, mutant enzyme R243K Pseudomonas aeruginosa
6.9
-
L-arginine pH 5.6, 25°C, mutant R243K Pseudomonas aeruginosa
8.7
-
L-Arg pH 5.6, 25°C, mutant enzyme R401A Pseudomonas aeruginosa
8.7
-
L-arginine pH 5.6, 25°C, mutant R401A Pseudomonas aeruginosa
15
-
L-Arg pH 5.6, 25°C, mutant enzyme R185A Pseudomonas aeruginosa
15
-
L-arginine pH 5.6, 25°C, mutant R185A Pseudomonas aeruginosa
21
-
Nomega-methyl-Arg pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
21
-
Nomega-methyl-L-arginine pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
38
-
L-Arg pH 5.6, 25°C, mutant enzyme R243L Pseudomonas aeruginosa
38
-
L-arginine pH 5.6, 25°C, mutant R243L Pseudomonas aeruginosa
50
-
Nomega-amino-L-arginine pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
50
-
Nomega-amino-Arg pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa

Metals/Ions

Metals/Ions Comment Organism Structure
KCl increases the activity 6fold at 60 mM Pseudomonas aeruginosa
MgCl2 increases the activity 8fold at 20 mM Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arginine + H2O Pseudomonas aeruginosa ADI catalyzes the first step of the energy-producing arginine degradation pathway found in certain microorganisms, which ultimately leads to ornithine, carbamate, and ATP, overview L-citrulline + NH3
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa
-
-
-
Pseudomonas aeruginosa P13981
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzyme Pseudomonas aeruginosa
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Pseudomonas aeruginosa

Reaction

Reaction Comment Organism Reaction ID
L-arginine + H2O = L-citrulline + NH3 residues Cys406, His278, and Asp166 and the contribution from the neighboring Asp280 are important in catalysis of the formation and hydrolysis of the Cys406-alkyluronium intermediate, structure and kinetic analysis Pseudomonas aeruginosa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Arg + H2O
-
Pseudomonas aeruginosa L-citrulline + NH3
-
?
L-arginine + H2O specific for Pseudomonas aeruginosa L-citrulline + NH3
-
?
L-arginine + H2O ADI catalyzes the first step of the energy-producing arginine degradation pathway found in certain microorganisms, which ultimately leads to ornithine, carbamate, and ATP, overview Pseudomonas aeruginosa L-citrulline + NH3
-
?
additional information no activity with agmatine Pseudomonas aeruginosa ?
-
?
Nomega-amino-Arg + H2O
-
Pseudomonas aeruginosa L-citrulline + hydrazine
-
?
Nomega-amino-L-arginine + H2O
-
Pseudomonas aeruginosa L-citrulline + hydrazine
-
?
Nomega-methyl-Arg + H2O
-
Pseudomonas aeruginosa L-citrulline + methylamine
-
?
Nomega-methyl-L-arginine + H2O
-
Pseudomonas aeruginosa L-citrulline + methylamine
-
?

Subunits

Subunits Comment Organism
More ADI C406A-L-arginine structure, overview Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
ADI
-
Pseudomonas aeruginosa
More ADI belongs to the guanidino group-modifying enzyme superfamily, GMSF, which conserves backbone fold and a Cys-, His-, and Asp-based catalytic core Pseudomonas aeruginosa
PaADI
-
Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Pseudomonas aeruginosa

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
additional information
-
no activity at 45°C Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00036
-
L-Arg pH 5.6, 25°C, mutant enzyme R401L Pseudomonas aeruginosa
0.00089
-
L-Arg pH 5.6, 25°C, mutant enzyme R243A Pseudomonas aeruginosa
0.002
-
L-Arg pH 5.6, 25°C, mutant enzyme R243K Pseudomonas aeruginosa
0.0021
-
L-Arg pH 5.6, 25°C, mutant enzyme E224A Pseudomonas aeruginosa
0.0037
-
L-Arg pH 5.6, 25°C, mutant enzyme E224D Pseudomonas aeruginosa
0.0072
-
L-Arg pH 5.6, 25°C, mutant enzyme R401A Pseudomonas aeruginosa
0.0084
-
L-Arg pH 5.6, 25°C, mutant enzyme R185L Pseudomonas aeruginosa
0.009
-
L-Arg pH 5.6, 25°C, mutant enzyme R243L Pseudomonas aeruginosa
0.01
-
Nomega-methyl-Arg pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
0.021
-
L-Arg pH 5.6, 25°C, mutant enzyme N160A Pseudomonas aeruginosa
0.089
-
L-Arg pH 5.6, 25°C, mutant enzyme R185A Pseudomonas aeruginosa
0.2
-
L-arginine pH 5.6, 25°C, mutant R243L Pseudomonas aeruginosa
0.3
-
L-arginine pH 5.6, 25°C, mutant R243A Pseudomonas aeruginosa
0.3
-
L-arginine pH 5.6, 25°C, mutant R243K Pseudomonas aeruginosa
0.8
-
L-arginine pH 5.6, 25°C, mutant R401A Pseudomonas aeruginosa
0.9
-
L-arginine pH 5.6, 25°C, mutant E224D Pseudomonas aeruginosa
1
-
Nomega-methyl-L-arginine pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
1.8
-
L-arginine pH 5.6, 25°C, mutant R185L Pseudomonas aeruginosa
1.9
-
L-arginine pH 5.6, 25°C, mutant E224A Pseudomonas aeruginosa
5.9
-
L-arginine pH 5.6, 25°C, mutant R185A Pseudomonas aeruginosa
6.3
-
L-arginine pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
6.3
-
L-Arg pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
7
-
Nomega-amino-L-arginine pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
7
-
Nomega-amino-Arg pH 5.6, 25°C, wild-type enzyme Pseudomonas aeruginosa
8
-
L-arginine pH 5.6, 25°C, mutant N160A Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.6
-
assay at Pseudomonas aeruginosa

pH Range

pH Minimum pH Maximum Comment Organism
5 8.6 pH profiles of wild-type and mutant enzymes Pseudomonas aeruginosa