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Literature summary for 2.3.1.5 extracted from

  • Kubiak, X.; Li de la Sierra-Gallay, I.; Chaffotte, A.F.; Pluvinage, B.; Weber, P.; Haouz, A.; Dupret, J.M.; Rodrigues-Lima, F.
    Structural and biochemical characterization of an active arylamine N-acetyltransferase possessing a non-canonical Cys-His-Glu catalytic triad (2013), J. Biol. Chem., 288, 22493-22505.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 cells Bacillus cereus

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 1.6 M sodium citrate, pH 6.5, 0.28 M NDSB-221 as additive Bacillus cereus

Protein Variants

Protein Variants Comment Organism
E123D the catalytic efficiency is 1.6-4.4 times higher than that of the wild type enzyme Bacillus cereus

Inhibitors

Inhibitors Comment Organism Structure
iodoacetamide
-
Bacillus cereus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
acetyl-CoA wild type enzyme, apparent value, with 4-aminosalicylic acid as cosubstrate, at pH 7.5 and 25°C Bacillus cereus
0.052
-
acetyl-CoA mutant enzyme E123D, apparent value, with 4-aminosalicylic acid as cosubstrate, at pH 7.5 and 25°C Bacillus cereus
0.64
-
4-aminosalicylic acid mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
0.86
-
2-Aminofluorene mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
1.34
-
4-aminosalicylic acid wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
5.8
-
2-Aminofluorene wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
14.55
-
isoniazid mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
76.9
-
isoniazid wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33465
-
x * 33465, calculated from amino acid sequence Bacillus cereus
34000
-
x * 34000, SDS-PAGE Bacillus cereus

Organism

Organism UniProt Comment Textmining
Bacillus cereus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Mono Q column chromatography Bacillus cereus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + 2-aminofluorene
-
Bacillus cereus CoA + N-acetyl-2-aminofluorene
-
?
acetyl-CoA + 4-aminosalicylic acid
-
Bacillus cereus CoA + N-acetyl-4-aminosalicylate
-
?
acetyl-CoA + isoniazid
-
Bacillus cereus CoA + N-acetylisoniazid
-
?

Subunits

Subunits Comment Organism
? x * 34000, SDS-PAGE Bacillus cereus
? x * 33465, calculated from amino acid sequence Bacillus cereus

Synonyms

Synonyms Comment Organism
NAT3
-
Bacillus cereus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
14.3
-
acetyl-CoA mutant enzyme E123D, apparent value, with 4-aminosalicylic acid as cosubstrate, at pH 7.5 and 25°C Bacillus cereus
19.3
-
acetyl-CoA wild type enzyme, apparent value, with 4-aminosalicylic acid as cosubstrate, at pH 7.5 and 25°C Bacillus cereus
43.9
-
4-aminosalicylic acid mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
44.1
-
2-Aminofluorene mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
56.6
-
4-aminosalicylic acid wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
67.4
-
2-Aminofluorene wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
167.1
-
isoniazid mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
289.3
-
isoniazid wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.76
-
isoniazid wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
11.5
-
isoniazid mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
11.5
-
2-Aminofluorene wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
42.4
-
4-aminosalicylic acid wild type enzyme, apparent value, at pH 7.5 and 25°C Bacillus cereus
51.3
-
2-Aminofluorene mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
68.4
-
4-aminosalicylic acid mutant enzyme E123D, apparent value, at pH 7.5 and 25°C Bacillus cereus
275
-
acetyl-CoA mutant enzyme E123D, apparent value, with 4-aminosalicylic acid as cosubstrate, at pH 7.5 and 25°C Bacillus cereus
390
-
acetyl-CoA wild type enzyme, apparent value, with 4-aminosalicylic acid as cosubstrate, at pH 7.5 and 25°C Bacillus cereus