Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Tris-HCl | activation at 0.25 M, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. |
General Stability | Organism |
---|---|
purified enzyme is very stable, considerably less stable in Tris-HCl than in phosphate buffer | Clostridium sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0034 | - |
acetoacetyl-CoA | pH 7.0, 25°C, cosubstrate crotonate | Clostridium sp. | |
0.023 | - |
butyryl-CoA | pH 7.0, 25°C, cosubstrate crotonate | Clostridium sp. | |
0.034 | - |
butyryl-CoA | pH 7.0, 25°C, cosubstrate acetoacetate | Clostridium sp. | |
0.044 | - |
butyryl-CoA | pH 6.3, 25°C, cosubstrate acetoacetate | Clostridium sp. | |
0.054 | - |
butyryl-CoA | pH 8.1, 25°C, cosubstrate acetoacetate 1.0 mM, 20 mM MgCl2 | Clostridium sp. | |
0.073 | - |
butyryl-CoA | pH 8.1, 25°C, cosubstrate acetoacetate 2.0 mM, 20 mM MgCl2 | Clostridium sp. | |
0.083 | - |
butyryl-CoA | pH 8.1, 25°C, cosubstrate acetoacetate 3.0 mM, 20 mM MgCl2 | Clostridium sp. | |
0.135 | - |
acetyl-CoA | pH 7.0, 25°C, cosubstrate acetoacetate | Clostridium sp. | |
0.167 | - |
crotonyl-CoA | pH 7.0, 25°C, cosubstrate acetoacetate | Clostridium sp. | |
0.205 | - |
propionyl-CoA | pH 7.0, 25°C, cosubstrate acetoacetate | Clostridium sp. | |
0.8 | - |
Butanoate | pH 7.5, 25°C | Clostridium sp. | |
1.1 | - |
acetoacetate | pH 7.5, 25°C, butyryl-CoA level of 0.18 mM | Clostridium sp. | |
1.4 | - |
acetoacetate | pH 7.5, 25°C, butyryl-CoA level of 0.23 mM | Clostridium sp. | |
1.8 | - |
acetoacetate | pH 7.5, 25°C, butyryl-CoA level of 0.37 mM | Clostridium sp. | |
2 | - |
crotonate | pH 7.5, 25°C | Clostridium sp. | |
3.2 | - |
propionate | pH 7.5, 25°C | Clostridium sp. | |
7 | - |
pentanoate | pH 7.5, 25°C | Clostridium sp. | |
14 | - |
Isobutyrate | pH 7.5, 25°C | Clostridium sp. | |
17 | - |
Vinylacetate | pH 7.5, 25°C | Clostridium sp. | |
21 | - |
Isopentanoate | pH 7.5, 25°C | Clostridium sp. | |
21 | - |
DL-2-Hydroxypentanoate | pH 7.5, 25°C | Clostridium sp. | |
24 | - |
acetate | pH 7.5, 25°C | Clostridium sp. | |
29 | - |
3-Mercaptopropionate | pH 7.5, 25°C | Clostridium sp. | |
29 | - |
Monochloroacetate | pH 7.5, 25°C | Clostridium sp. | |
33 | - |
4-hydroxybuyrate | pH 7.5, 25°C | Clostridium sp. | |
40 | - |
Acrylate | pH 7.5, 25°C | Clostridium sp. | |
42 | - |
3-Methylvinylacetate | pH 7.5, 25°C | Clostridium sp. | |
44 | - |
DL-3-hydroxybutyrate | pH 7.5, 25°C | Clostridium sp. | |
49 | - |
DL-2-hydroxybuyrate | pH 7.5, 25°C | Clostridium sp. | |
250 | - |
3-hydroxypropionate | pH 7.5, 25°C | Clostridium sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activation at 6 mM CaCl2, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. | |
K+ | activation at 0.08 M KCl, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. | |
Li+ | activation at 0.2 M LiCl, inhibitions at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. | |
Mg2+ | activation at 12 mM MgCl2, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. | |
Mn2+ | activation at 1.2 mM MnCl2, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. | |
Na+ | activation at 0.17 M NaCl, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. | |
NH4+ | activation at 0.08 M NH4Cl, inhibition at higher concentrations, only with acetoacetate or acetoacetyl-CoA as substrate | Clostridium sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
23300 | - |
2 * 23300 + 2 * 25100, 2 alpha and 2 beta subunits, SDS-PAGE | Clostridium sp. |
25100 | - |
2 * 23300 + 2 * 25100, 2 alpha and 2 beta subunits, SDS-PAGE | Clostridium sp. |
90000 | - |
gel electrophoresis | Clostridium sp. |
108000 | - |
gel filtration | Clostridium sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
butanoate + acetoacetyl-CoA | Acetoanaerobium sticklandii | - |
butanoyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | Clostridium sp. | lysine degradation pathway | butanoyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | Clostridium sp. SB4 | lysine degradation pathway | butanoyl-CoA + acetoacetate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetoanaerobium sticklandii | - |
- |
- |
Clostridium sp. | - |
SB4 | - |
Clostridium sp. SB4 | - |
SB4 | - |
Purification (Comment) | Organism |
---|---|
- |
Clostridium sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
329 | - |
- |
Clostridium sp. |
Storage Stability | Organism |
---|---|
-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. |
-15°C, 3.1 mg protein/ml, 20 mM potassium phosphate, pH 7.0, retains full activity during storage for 1 year | Clostridium sp. |
4°C, 0.0062 mg/ml protein, 10 mM phosphate buffer, pH 7.0, half-life of about 7 days | Clostridium sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxypropanoate + acetoacetyl-CoA | reaction at 8.6% the rate of butyrate | Clostridium sp. | 3-hydroxypropanoyl-CoA + acetoacetate | - |
r | |
3-mercaptopropionate + acetoacetyl-CoA | reaction at 71% the rate of butyrate | Clostridium sp. | 3-mercaptopropionyl-CoA + acetoacetate | - |
r | |
3-methylvinylacetate + acetoacetyl-CoA | reaction at 44% the rate of butyrate | Clostridium sp. | 3-methylvinylacetyl-CoA + acetoacetate | - |
r | |
4-hydroxybutyrate + acetoacetyl-CoA | reaction at 14% the rate of butyrate | Clostridium sp. | 4-hydroxybutanoyl-CoA + acetoacetate | - |
r | |
acetate + acetoacetyl-CoA | - |
Clostridium sp. | acetyl-CoA + acetoacetate | - |
r | |
acetate + acetoacetyl-CoA | reaction at 24% the rate of butyrate | Clostridium sp. | acetyl-CoA + acetoacetate | - |
r | |
acetate + acetoacetyl-CoA | - |
Clostridium sp. SB4 | acetyl-CoA + acetoacetate | - |
r | |
acetate + acetoacetyl-CoA | reaction at 24% the rate of butyrate | Clostridium sp. SB4 | acetyl-CoA + acetoacetate | - |
r | |
acrylate + acetoacetyl-CoA | reaction at 10% the rate of butyrate | Clostridium sp. | acrylyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | - |
Acetoanaerobium sticklandii | butanoyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | - |
Clostridium sp. | butanoyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | lysine degradation pathway | Clostridium sp. | butanoyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | - |
Clostridium sp. SB4 | butanoyl-CoA + acetoacetate | - |
r | |
butanoate + acetoacetyl-CoA | lysine degradation pathway | Clostridium sp. SB4 | butanoyl-CoA + acetoacetate | - |
r | |
butanoyl-CoA + 2-methylacetoacetate | used 12% as fast as acetoacetate | Clostridium sp. | butanoate + 2-methylacetoacetyl-CoA | - |
r | |
butanoyl-CoA + 3-oxohexanoate | used 21% as fast as acetoacetate | Clostridium sp. | butanoate + 3-oxohexanoyl-CoA | - |
r | |
butanoyl-CoA + 3-oxopentanoate | used 60% as fast as acetoacetate | Clostridium sp. | butanoate + 3-oxopentanoyl-CoA | - |
r | |
butanoyl-CoA + acetoacetate | - |
Clostridium sp. | butanoate + acetoacetyl-CoA | - |
r | |
crotonate + acetoacetyl-CoA | reaction at 16% the rate of butyrate | Clostridium sp. | crotonyl-CoA + acetoacetate | - |
r | |
DL-2-hydroxybutyrate + acetoacetyl-CoA | reaction at 19% the rate of butyrate | Clostridium sp. | 2-hydroxybutyryl-CoA + acetoacetate | - |
r | |
DL-2-hydroxypentanoate + acetoacetyl-CoA | reaction at 7.9% the rate of butyrate | Clostridium sp. | 2-hydroxypentanoyl-CoA + acetoacetate | - |
r | |
DL-3-hydroxybutyrate + acetoacetyl-CoA | reaction at 30% the rate of butyrate | Clostridium sp. | 3-hydroxybutyryl-CoA + acetoacetate | - |
r | |
isobutyrate + acetoacetyl-CoA | reaction at 19% the rate of butyrate | Clostridium sp. | isobutyryl-CoA + acetoacetate | - |
r | |
isopentanoate + acetoacetyl-CoA | reaction at 8.3% the rate of butyrate | Clostridium sp. | isopentanoyl-CoA + acetoacetate | - |
r | |
monochloroacetate + acetoacetyl-CoA | reaction at 5.4% the rate of butyrate | Clostridium sp. | monochloroacetyl-CoA + acetoacetate | - |
r | |
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. | ? | - |
? | |
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 | ? | - |
? | |
pentanoate + acetoacetyl-CoA | reaction at 19% the rate of butyrate | Clostridium sp. | pentanoyl-CoA + acetoacetate | - |
r | |
propionate + acetoacetyl-CoA | reaction at 21% the rate of butyrate | Clostridium sp. | propionyl-CoA + acetoacetate | - |
r | |
vinylacetate + acetoacetyl-CoA | best substrate | Clostridium sp. | vinylacetyl-CoA + acetoacetate | - |
r |
Subunits | Comment | Organism |
---|---|---|
tetramer | 2 * 23300 + 2 * 25100, 2 alpha and 2 beta subunits, SDS-PAGE | Clostridium sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8 | - |
Clostridium sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 9 | maximally active from pH 6.6 to pH 7.9 | Clostridium sp. |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6.5 | 7 | 1 h, at 30°C, most stable, about 20% loss of activity at pH 6.0 and 8.0 | Clostridium sp. |