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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 0.0034
-
 acetoacetyl-CoA pH 7.0, 25°C, cosubstrate crotonate Clostridium sp.
2.8.3.9 0.023
-
 butyryl-CoA pH 7.0, 25°C, cosubstrate crotonate Clostridium sp.
2.8.3.9 0.034
-
 butyryl-CoA pH 7.0, 25°C, cosubstrate acetoacetate Clostridium sp.
2.8.3.9 0.044
-
 butyryl-CoA pH 6.3, 25°C, cosubstrate acetoacetate 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 0.073
-
 butyryl-CoA pH 8.1, 25°C, cosubstrate acetoacetate 2.0 mM, 20 mM MgCl2 Clostridium sp.
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 0.135
-
 acetyl-CoA pH 7.0, 25°C, cosubstrate acetoacetate Clostridium sp.
2.8.3.9 0.167
-
 crotonyl-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.
2.8.3.9 0.8
-
 Butanoate 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 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 2
-
 crotonate 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 7
-
 pentanoate pH 7.5, 25°C Clostridium sp.
2.8.3.9 14
-
 Isobutyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 17
-
 Vinylacetate 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 24
-
 acetate pH 7.5, 25°C 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 33
-
 4-hydroxybuyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 40
-
 Acrylate pH 7.5, 25°C Clostridium sp.
2.8.3.9 42
-
 3-Methylvinylacetate 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 49
-
 DL-2-hydroxybuyrate pH 7.5, 25°C Clostridium sp.
2.8.3.9 250
-
 3-hydroxypropionate pH 7.5, 25°C Clostridium sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
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 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 Li+ activation at 0.2 M LiCl, inhibitions 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 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 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 NH4+ activation at 0.08 M NH4Cl, inhibition 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 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.
2.8.3.9 90000
-
 gel electrophoresis Clostridium sp.
2.8.3.9 108000
-
 gel filtration 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 Acetoanaerobium sticklandii
-
-
-
2.8.3.9 Clostridium sp.
-
 SB4
-
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 -15°C, 3.1 mg protein/ml, 20 mM potassium phosphate, pH 7.0, retains full activity during storage for 1 year 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.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 3-mercaptopropionate + acetoacetyl-CoA reaction at 71% the rate of butyrate Clostridium sp. 3-mercaptopropionyl-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 4-hydroxybutyrate + acetoacetyl-CoA reaction at 14% the rate of butyrate Clostridium sp. 4-hydroxybutanoyl-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 acrylate + acetoacetyl-CoA reaction at 10% the rate of butyrate Clostridium sp. acrylyl-CoA + acetoacetate
-
 r
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 butanoyl-CoA + 2-methylacetoacetate used 12% as fast as acetoacetate Clostridium sp. butanoate + 2-methylacetoacetyl-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 + 3-oxopentanoate used 60% as fast as acetoacetate Clostridium sp. butanoate + 3-oxopentanoyl-CoA
-
 r
2.8.3.9 butanoyl-CoA + acetoacetate
-
 Clostridium sp. butanoate + acetoacetyl-CoA
-
 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 DL-2-hydroxybutyrate + acetoacetyl-CoA reaction at 19% the rate of butyrate Clostridium sp. 2-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 DL-3-hydroxybutyrate + acetoacetyl-CoA reaction at 30% the rate of butyrate Clostridium sp. 3-hydroxybutyryl-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 isopentanoate + acetoacetyl-CoA reaction at 8.3% the rate of butyrate Clostridium sp. isopentanoyl-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 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 pentanoate + acetoacetyl-CoA reaction at 19% the rate of butyrate Clostridium sp. pentanoyl-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 vinylacetate + acetoacetyl-CoA best substrate Clostridium sp. vinylacetyl-CoA + acetoacetate
-
 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.