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Literature summary for 3.5.1.4 extracted from

  • Gregoriou, M.; Brown, P.R.
    Inhibition of the aliphatic amidase from Pseudomonas aeruginosa by urea and related compounds (1979), Eur. J. Biochem., 96, 101-108.
    View publication on PubMed
Search for more literature for 3.5.1.4

Inhibitors

Inhibitors Comment Organism Structure
acetaldehyde competitive inhibitor, not time dependent; competitve inhibitor, Ki: 7.0 micromol Pseudomonas aeruginosa
acetaldehyde ammonia
-
 Pseudomonas aeruginosa
Cyanate Ki of strain AI3 is 1.1 mM, Ki of strain AIU1N is 1.8 mM and Ki of strain OUCH4 is 50 mM; reversible inhibition, pH 7.2 and 37°C Pseudomonas aeruginosa
Hydroxyurea Ki of strain AI3 is 1.4 mM, Ki of strain AIU1N is 10.8 mM and Ki of strain OUCH4 is 337 mM, regaines full activity within 24 h at 37°C in 0.05 M Tris buffer at pH 7.2; reversible inhibition, pH 7.2 and 37°C Pseudomonas aeruginosa
additional information no inhibition with 10 mM guanidinium carbonate, 100 mM thiourea, ammonium carbamate at strain AI 3 Pseudomonas aeruginosa
Urea competitive inhibitor at pH 7.2, 90 mM acetamide protectes AI3 amidase against 2.9 mM urea over 2 h, Ki of strain AI3 is 6.6 mM, Ki of strain AIU1N is 41.3 mM and Ki of strain OUCH4 is not known, regaines full activity within 24 h at 37°C in 0.05 M Tris buffer at pH 7.2; reversible inhibition, pH 7.2 and 37°C Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
68.6
-
 acetamide strain AI3 Pseudomonas aeruginosa
90.4
-
 acetamide strain AIU1N Pseudomonas aeruginosa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
41600
-
 6 * 41600, SDS-PAGE Pseudomonas aeruginosa
240000
-
 SDS-PAGE Pseudomonas aeruginosa
240000
-
 calculated from measurement of the subunit molecular weight, strain AI3 Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetanilide + H2O Pseudomonas aeruginosa
-
 acetate + aniline
-
 ?
acetanilide + H2O Pseudomonas aeruginosa AI3
-
 acetate + aniline
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa
-
 and the mutant strains AIU1N and OUCH4
-
Pseudomonas aeruginosa
-
 and strains AIU1N and OUCH4
-
Pseudomonas aeruginosa AI3
-
 and the mutant strains AIU1N and OUCH4
-
Pseudomonas aeruginosa AI3
-
 and strains AIU1N and OUCH4
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.8
-
 at 3 mM formamide, formamidase activity of AI3 amidase Pseudomonas aeruginosa
1.5
-
 substrate 4-nitro-acetanilide, strain OUCH4 Pseudomonas aeruginosa
1.5
-
 4-nitroacetanilide, strain OUCH 4, pH 8.5 Pseudomonas aeruginosa
7.4
-
 substrate 4-nitro-acetanilide, strain AIU1N Pseudomonas aeruginosa
7.4
-
 4-nitroacetanilide, strain AIU 1N, pH 8.5 Pseudomonas aeruginosa
10.5
-
 substrate 4-nitro-acetanilide, strain AI3 25-fold greater activity than wild-type Pseudomonas aeruginosa
10.5
-
 4-nitroacetanilide, strain AI3, pH 8.5 Pseudomonas aeruginosa
2900
-
 substrate acetamide, hydrolysis, strain AIU1N Pseudomonas aeruginosa
4100
-
 substrate acetamide, hydrolysis, strain AI3 Pseudomonas aeruginosa

Storage Stability

Storage Stability Organism
-30°C, 0.05 M Tris buffer, pH 7.2, 50% v/v glycerol, 1 mM 2-mercaptoethanol Pseudomonas aeruginosa
purified amidase stored in 0.05 M Tris buffer, pH 7.2, containing 1 mM mercaptoethanol and 50%, v/v, glycerol, at -30°C Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitroacetanilide + H2O
-
 Pseudomonas aeruginosa acetic acid + 4-nitroaniline
-
 ?
4-nitroacetanilide + H2O
-
 Pseudomonas aeruginosa AI3 acetic acid + 4-nitroaniline
-
 ?
acetamide + H2O
-
 Pseudomonas aeruginosa acetic acid + NH3
-
 ?
acetamide + H2O
-
 Pseudomonas aeruginosa AI3 acetic acid + NH3
-
 ?
acetanilide + H2O
-
 Pseudomonas aeruginosa acetate + aniline
-
 ?
acetanilide + H2O
-
 Pseudomonas aeruginosa AI3 acetate + aniline
-
 ?

Subunits

Subunits Comment Organism
hexamer 6 * 41600, SDS-PAGE Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.143
-
 acetamide hydrolysis, strain AI3 Pseudomonas aeruginosa
1.51
-
 acetamide hydrolysis, strain AIU1N Pseudomonas aeruginosa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.005
-
 acetaldehyde strain AI3 Pseudomonas aeruginosa
0.0051
-
 acetaldehyde competitive inhibitor Pseudomonas aeruginosa
0.007
-
 acetaldehyde ammonia strain AI3 Pseudomonas aeruginosa
1.1
-
 Cyanate strain AI3, pH 7.2 and 37°C Pseudomonas aeruginosa
1.4
-
 Hydroxyurea strain AI3, pH 7.2 and 37°C Pseudomonas aeruginosa
1.8
-
 Cyanate strain AIU1N, pH 7.2 and 37°C Pseudomonas aeruginosa
6.6
-
 Urea strain AI3, pH 7.2 and 37°C Pseudomonas aeruginosa
10.8
-
 Hydroxyurea strain AIU1N, pH 7.2 and 37°C Pseudomonas aeruginosa
41.3
-
 Urea strain AIU1N pH 7.2 and 37°C Pseudomonas aeruginosa
50
-
 Cyanate strain OUCH4, pH 7.2 and 37°C Pseudomonas aeruginosa
337
-
 Hydroxyurea strain OUCH4, pH 7.2 and 37°C Pseudomonas aeruginosa