Literature summary extracted from

  • Barker, H.A.; Jemg, I.M.; Neff, N.; Robertson, J.M.; Tam, F.K.; Hosaka, S.
    Butyryl-CoA:acetoacetate CoA-transferase from a lysine-fermenting Clostridium (1978), J. Biol. Chem., 253, 1219-1225.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.8.3.9 Tris-HCl activation at 0.25 M, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.

General Stability

EC Number General Stability Organism
2.8.3.9 purified enzyme is very stable, considerably less stable in Tris-HCl than in phosphate buffer Clostridium sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.8.3.9 0.083
-
butyryl-CoA pH 8.1, 25°C, cosubstrate acetoacetate 3.0 mM, 20 mM MgCl2 Clostridium sp.
2.8.3.9 7
-
pentanoate pH 7.5, 25°C Clostridium sp.
2.8.3.9 2
-
crotonate pH 7.5, 25°C Clostridium sp.
2.8.3.9 0.023
-
butyryl-CoA pH 7.0, 25°C, cosubstrate crotonate Clostridium sp.
2.8.3.9 24
-
acetate pH 7.5, 25°C Clostridium sp.
2.8.3.9 0.8
-
Butanoate pH 7.5, 25°C Clostridium sp.
2.8.3.9 1.4
-
acetoacetate pH 7.5, 25°C, butyryl-CoA level of 0.23 mM Clostridium sp.
2.8.3.9 1.8
-
acetoacetate pH 7.5, 25°C, butyryl-CoA level of 0.37 mM Clostridium sp.
2.8.3.9 0.034
-
butyryl-CoA pH 7.0, 25°C, cosubstrate acetoacetate Clostridium sp.
2.8.3.9 17
-
Vinylacetate pH 7.5, 25°C Clostridium sp.
2.8.3.9 1.1
-
acetoacetate pH 7.5, 25°C, butyryl-CoA level of 0.18 mM Clostridium sp.
2.8.3.9 0.0034
-
acetoacetyl-CoA pH 7.0, 25°C, cosubstrate crotonate Clostridium sp.
2.8.3.9 0.073
-
butyryl-CoA pH 8.1, 25°C, cosubstrate acetoacetate 2.0 mM, 20 mM MgCl2 Clostridium sp.
2.8.3.9 14
-
Isobutyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 40
-
Acrylate pH 7.5, 25°C Clostridium sp.
2.8.3.9 0.054
-
butyryl-CoA pH 8.1, 25°C, cosubstrate acetoacetate 1.0 mM, 20 mM MgCl2 Clostridium sp.
2.8.3.9 33
-
4-hydroxybuyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 0.044
-
butyryl-CoA pH 6.3, 25°C, cosubstrate acetoacetate Clostridium sp.
2.8.3.9 29
-
3-Mercaptopropionate pH 7.5, 25°C Clostridium sp.
2.8.3.9 29
-
Monochloroacetate pH 7.5, 25°C Clostridium sp.
2.8.3.9 0.167
-
crotonyl-CoA pH 7.0, 25°C, cosubstrate acetoacetate Clostridium sp.
2.8.3.9 42
-
3-Methylvinylacetate pH 7.5, 25°C Clostridium sp.
2.8.3.9 49
-
DL-2-hydroxybuyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 3.2
-
propionate pH 7.5, 25°C Clostridium sp.
2.8.3.9 250
-
3-hydroxypropionate pH 7.5, 25°C Clostridium sp.
2.8.3.9 44
-
DL-3-hydroxybutyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 21
-
Isopentanoate pH 7.5, 25°C Clostridium sp.
2.8.3.9 21
-
DL-2-Hydroxypentanoate pH 7.5, 25°C Clostridium sp.
2.8.3.9 0.135
-
acetyl-CoA pH 7.0, 25°C, cosubstrate acetoacetate Clostridium sp.
2.8.3.9 0.205
-
propionyl-CoA pH 7.0, 25°C, cosubstrate acetoacetate Clostridium sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.8.3.9 Mn2+ activation at 1.2 mM MnCl2, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.
2.8.3.9 Ca2+ activation at 6 mM CaCl2, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.
2.8.3.9 Mg2+ activation at 12 mM MgCl2, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.
2.8.3.9 K+ activation at 0.08 M KCl, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.
2.8.3.9 NH4+ activation at 0.08 M NH4Cl, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.
2.8.3.9 Na+ activation at 0.17 M NaCl, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.
2.8.3.9 Li+ activation at 0.2 M LiCl, inhibitions at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate Clostridium sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.8.3.9 90000
-
gel electrophoresis Clostridium sp.
2.8.3.9 108000
-
gel filtration Clostridium sp.
2.8.3.9 23300
-
2 * 23300 + 2 * 25100, 2 alpha and 2 beta subunits, SDS-PAGE Clostridium sp.
2.8.3.9 25100
-
2 * 23300 + 2 * 25100, 2 alpha and 2 beta subunits, SDS-PAGE Clostridium sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.8.3.9 butanoate + acetoacetyl-CoA Acetoanaerobium sticklandii
-
butanoyl-CoA + acetoacetate
-
r
2.8.3.9 butanoate + acetoacetyl-CoA Clostridium sp. lysine degradation pathway butanoyl-CoA + acetoacetate
-
r
2.8.3.9 butanoate + acetoacetyl-CoA Clostridium sp. SB4 lysine degradation pathway butanoyl-CoA + acetoacetate
-
r

Organism

EC Number Organism UniProt Comment Textmining
2.8.3.9 Clostridium sp.
-
SB4
-
2.8.3.9 Acetoanaerobium sticklandii
-
-
-
2.8.3.9 Clostridium sp. SB4
-
SB4
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.8.3.9
-
Clostridium sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.8.3.9 329
-
-
Clostridium sp.

Storage Stability

EC Number Storage Stability Organism
2.8.3.9 -15°C 0.0062 mg/ml protein, 10 mM phosphate buffer, pH 7.0, half-life of 12 days when thawed and refrozen 5 times Clostridium sp.
2.8.3.9 4°C, 0.0062 mg/ml protein, 10 mM phosphate buffer, pH 7.0, half-life of about 7 days Clostridium sp.
2.8.3.9 -15°C, 3.1 mg protein/ml, 20 mM potassium phosphate, pH 7.0, retains full activity during storage for 1 year Clostridium sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.8.3.9 additional information monofluoroacetate and hexanoate are no substrates, compounds found to be unreactive or less than 0.1% as reactive as butyrate are formate, cyanoacetate, n-heptanoate, tiglate, oxalate, malonate, succinate, citramalate, DL-alpha-hydroxyisobutyrate, 3,4-dihydroxybutyrate, alpha-ketobutyrate, 2-oxoglutarate, DL-3-aminobutyrate and 4-aminobutyrate, 3-oxo-5-aminohexanoate and triacetate are unreactive Clostridium sp. ?
-
?
2.8.3.9 additional information monofluoroacetate and hexanoate are no substrates, compounds found to be unreactive or less than 0.1% as reactive as butyrate are formate, cyanoacetate, n-heptanoate, tiglate, oxalate, malonate, succinate, citramalate, DL-alpha-hydroxyisobutyrate, 3,4-dihydroxybutyrate, alpha-ketobutyrate, 2-oxoglutarate, DL-3-aminobutyrate and 4-aminobutyrate, 3-oxo-5-aminohexanoate and triacetate are unreactive Clostridium sp. SB4 ?
-
?
2.8.3.9 butanoate + acetoacetyl-CoA
-
Acetoanaerobium sticklandii butanoyl-CoA + acetoacetate
-
r
2.8.3.9 butanoate + acetoacetyl-CoA
-
Clostridium sp. butanoyl-CoA + acetoacetate
-
r
2.8.3.9 butanoate + acetoacetyl-CoA lysine degradation pathway Clostridium sp. butanoyl-CoA + acetoacetate
-
r
2.8.3.9 butanoate + acetoacetyl-CoA
-
Clostridium sp. SB4 butanoyl-CoA + acetoacetate
-
r
2.8.3.9 butanoate + acetoacetyl-CoA lysine degradation pathway Clostridium sp. SB4 butanoyl-CoA + acetoacetate
-
r
2.8.3.9 acetate + acetoacetyl-CoA
-
Clostridium sp. acetyl-CoA + acetoacetate
-
r
2.8.3.9 acetate + acetoacetyl-CoA reaction at 24% the rate of butyrate Clostridium sp. acetyl-CoA + acetoacetate
-
r
2.8.3.9 acetate + acetoacetyl-CoA
-
Clostridium sp. SB4 acetyl-CoA + acetoacetate
-
r
2.8.3.9 acetate + acetoacetyl-CoA reaction at 24% the rate of butyrate Clostridium sp. SB4 acetyl-CoA + acetoacetate
-
r
2.8.3.9 vinylacetate + acetoacetyl-CoA best substrate Clostridium sp. vinylacetyl-CoA + acetoacetate
-
r
2.8.3.9 propionate + acetoacetyl-CoA reaction at 21% the rate of butyrate Clostridium sp. propionyl-CoA + acetoacetate
-
r
2.8.3.9 3-mercaptopropionate + acetoacetyl-CoA reaction at 71% the rate of butyrate Clostridium sp. 3-mercaptopropionyl-CoA + acetoacetate
-
r
2.8.3.9 crotonate + acetoacetyl-CoA reaction at 16% the rate of butyrate Clostridium sp. crotonyl-CoA + acetoacetate
-
r
2.8.3.9 pentanoate + acetoacetyl-CoA reaction at 19% the rate of butyrate Clostridium sp. pentanoyl-CoA + acetoacetate
-
r
2.8.3.9 isobutyrate + acetoacetyl-CoA reaction at 19% the rate of butyrate Clostridium sp. isobutyryl-CoA + acetoacetate
-
r
2.8.3.9 3-methylvinylacetate + acetoacetyl-CoA reaction at 44% the rate of butyrate Clostridium sp. 3-methylvinylacetyl-CoA + acetoacetate
-
r
2.8.3.9 DL-3-hydroxybutyrate + acetoacetyl-CoA reaction at 30% the rate of butyrate Clostridium sp. 3-hydroxybutyryl-CoA + acetoacetate
-
r
2.8.3.9 DL-2-hydroxypentanoate + acetoacetyl-CoA reaction at 7.9% the rate of butyrate Clostridium sp. 2-hydroxypentanoyl-CoA + acetoacetate
-
r
2.8.3.9 isopentanoate + acetoacetyl-CoA reaction at 8.3% the rate of butyrate Clostridium sp. isopentanoyl-CoA + acetoacetate
-
r
2.8.3.9 acrylate + acetoacetyl-CoA reaction at 10% the rate of butyrate Clostridium sp. acrylyl-CoA + acetoacetate
-
r
2.8.3.9 monochloroacetate + acetoacetyl-CoA reaction at 5.4% the rate of butyrate Clostridium sp. monochloroacetyl-CoA + acetoacetate
-
r
2.8.3.9 4-hydroxybutyrate + acetoacetyl-CoA reaction at 14% the rate of butyrate Clostridium sp. 4-hydroxybutanoyl-CoA + acetoacetate
-
r
2.8.3.9 3-hydroxypropanoate + acetoacetyl-CoA reaction at 8.6% the rate of butyrate Clostridium sp. 3-hydroxypropanoyl-CoA + acetoacetate
-
r
2.8.3.9 DL-2-hydroxybutyrate + acetoacetyl-CoA reaction at 19% the rate of butyrate Clostridium sp. 2-hydroxybutyryl-CoA + acetoacetate
-
r
2.8.3.9 butanoyl-CoA + acetoacetate
-
Clostridium sp. butanoate + acetoacetyl-CoA
-
r
2.8.3.9 butanoyl-CoA + 3-oxopentanoate used 60% as fast as acetoacetate Clostridium sp. butanoate + 3-oxopentanoyl-CoA
-
r
2.8.3.9 butanoyl-CoA + 3-oxohexanoate used 21% as fast as acetoacetate Clostridium sp. butanoate + 3-oxohexanoyl-CoA
-
r
2.8.3.9 butanoyl-CoA + 2-methylacetoacetate used 12% as fast as acetoacetate Clostridium sp. butanoate + 2-methylacetoacetyl-CoA
-
r

Subunits

EC Number Subunits Comment Organism
2.8.3.9 tetramer 2 * 23300 + 2 * 25100, 2 alpha and 2 beta subunits, SDS-PAGE Clostridium sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.8.3.9 30
-
1 h, in potassium phosphate buffer, pH 6.5-7.0, most stable, about 20% loss of activity at pH 6.0 and 8.0 Clostridium sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.8.3.9 6.5 8
-
Clostridium sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.8.3.9 4 9 maximally active from pH 6.6 to pH 7.9 Clostridium sp.

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.8.3.9 6.5 7 1 h, at 30°C, most stable, about 20% loss of activity at pH 6.0 and 8.0 Clostridium sp.