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Literature summary for 4.1.1.4 extracted from

  • Ho, M.C.; Menetret, J.F.; Tsuruta, H.; Allen, K.N.
    The origin of the electrostatic perturbation in acetoacetate decarboxylase (2009), Nature, 459, 393-397.
    View publication on PubMed
Search for more literature for 4.1.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Clostridium acetobutylicum
expressed in Escherichia coli BL21(DE3) cells Chromobacterium violaceum

Crystallization (Commentary)

Crystallization (Comment) Organism
AAD complexed with 2,4-pentanedione, hanging drop vapor diffusion method, using 18-20% (v/v) glycerol, 40 mM phosphate buffer, pH 5.95, 100 mM sarcosine and 14-15% (w/v) PEG 3350 Clostridium acetobutylicum
AAD complexed with 2,4-pentanedione, hanging drop vapor diffusion method, using 18-20% (v/v) glycerol, 40 mM phosphate buffer, pH 5.95, 100 mM sarcosine and 14-15% (w/v) PEG 3350 Chromobacterium violaceum

Protein Variants

Protein Variants Comment Organism
E61Q the catalytic activity of the mutant shows a decrease in kcat (about 20fold with no change in Km) Clostridium acetobutylicum
E61Q the catalytic activity of the mutant shows a decrease in kcat (about 20fold with no change in Km) Chromobacterium violaceum
E76Q the catalytic activity of the mutant shows a decrease in kcat, the mutant exhibits a slight downward shift of the pH optimum Clostridium acetobutylicum
E76Q the catalytic activity of the mutant shows a decrease in kcat, the mutant exhibits aslight downward shift of the pH optimum Chromobacterium violaceum
R29Q the catalytic activity of Arg29Gln does not increase at pH values above the wild type optimum for AAD of about 5.4 Clostridium acetobutylicum
R29Q the catalytic activity of Arg29Gln does not increase at pH values above the wild type optimum for AAD of about 5.4 Chromobacterium violaceum

Inhibitors

Inhibitors Comment Organism Structure
2,4-Pentanedione potent inhibitor Chromobacterium violaceum
2,4-Pentanedione potent inhibitor Clostridium acetobutylicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1
-
 acetoacetate at 25°C, in 50 mM phosphate buffer, pH 5.95 Clostridium acetobutylicum
5.7
-
 acetoacetate at 25°C, in 50 mM phosphate buffer, pH 5.95 Chromobacterium violaceum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
365000
-
-
 Clostridium acetobutylicum
365000
-
-
 Chromobacterium violaceum

Organism

Organism UniProt Comment Textmining
Chromobacterium violaceum Q7NSA6
-
-
Clostridium acetobutylicum P23670
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and S-200 gel filtration Clostridium acetobutylicum
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and S-200 gel filtration Chromobacterium violaceum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-pentanedione
-
 Chromobacterium violaceum ?
-
 ?
acetoacetate + H+
-
 Clostridium acetobutylicum acetone + CO2
-
 ?
acetoacetate + H+
-
 Chromobacterium violaceum acetone + CO2
-
 ?

Subunits

Subunits Comment Organism
homodecamer x-ray crystallography Clostridium acetobutylicum
homodecamer x-ray crystallography Chromobacterium violaceum

Synonyms

Synonyms Comment Organism
AAD
-
 Clostridium acetobutylicum
AAD
-
 Chromobacterium violaceum
AADase
-
 Clostridium acetobutylicum
AADase
-
 Chromobacterium violaceum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
165
-
 acetoacetate at 25°C, in 50 mM phosphate buffer, pH 5.95 Clostridium acetobutylicum
349
-
 acetoacetate at 25°C, in 50 mM phosphate buffer, pH 5.95 Chromobacterium violaceum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4
-
-
 Clostridium acetobutylicum
5.4
-
-
 Chromobacterium violaceum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0007
-
 2,4-Pentanedione at 25°C, in 50 mM phosphate buffer, pH 5.95 Clostridium acetobutylicum