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Literature summary extracted from

  • Sedlmeier, H.; Simon, H.
    Purification and some properties of an acryloyl-CoA reductase of Clostridium kluyveri (1985), Biol. Chem. Hoppe-Seyler, 366, 953-961.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.1.95 ethyl acrylate strong inhibitor of the reduction of ethyl vinyl ketone Clostridium kluyveri
1.3.8.1 diethyl dicarbonate 1 mM, 83% residual activity Clostridium kluyveri
1.3.8.1 iodoacetamide 1 mM, 38% residual activity Clostridium kluyveri

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.95 0.00083
-
acryloyl-CoA in 0.1 M phosphate buffer pH 6.4, at 22°C Clostridium kluyveri
1.3.1.95 0.008
-
(E)-2-butenoyl-CoA in 0.1 M phosphate buffer pH 6.4, at 22°C Clostridium kluyveri
1.3.1.95 0.77
-
ethyl vinyl ketone in 0.1 M phosphate buffer pH 6.4, at 22°C Clostridium kluyveri
1.3.1.95 5.6
-
acrolein in 0.1 M phosphate buffer pH 6.4, at 22°C Clostridium kluyveri
1.3.1.95 35
-
N-acetyl-S-acryloylcysteamine in 0.1 M phosphate buffer pH 6.4, at 22°C Clostridium kluyveri
1.3.8.1 0.00083
-
acrylyl-CoA 22°C, pH 6.4 Clostridium kluyveri
1.3.8.1 0.008
-
(E)-2-butenoyl-CoA 22°C, pH 6.4 Clostridium kluyveri
1.3.8.1 0.035
-
N-acetyl-S-acryloyl-cysteamine 22°C, pH 6.4 Clostridium kluyveri
1.3.8.1 0.77
-
ethyl vinyl ketone 22°C, pH 6.4 Clostridium kluyveri
1.3.8.1 5.6
-
acrolein 22°C, pH 6.4 Clostridium kluyveri

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.3.1.95 additional information does not contain iron Clostridium kluyveri
1.3.8.1 Iron less than 0.2 g-atom per mol protein Clostridium kluyveri

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.1.95 14200
-
2 * 14200, SDS-PAGE Clostridium kluyveri
1.3.1.95 28400
-
gel filtration Clostridium kluyveri
1.3.8.1 14200
-
2 * 14200, SDS-PAGE Clostridium kluyveri
1.3.8.1 28400
-
gel filtration Clostridium kluyveri

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.95 Clostridium kluyveri
-
-
-
1.3.8.1 Clostridium kluyveri
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.3.1.95 the enzyme exhibits up to 11% and up to 16% residual activity after 19 days of exposure to air at pH 8.0 and 10°C and 30°C, respectively Clostridium kluyveri
1.3.8.1 enzyme is not oxygen-sensitive Clostridium kluyveri

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.95 DEAE-Sepharose CL-6B column chromatography, phenyl-Sepharose CL-4B column chromatography, CM-Sephadex A-25 gel filtration, and Sephacryl S-200 gel filtration Clostridium kluyveri

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.8.1 170
-
22°C, pH 6.4 Clostridium kluyveri

Storage Stability

EC Number Storage Stability Organism
1.3.1.95 -18°C, 30% (w/v) ethylene glycol in 15 mM phosphate buffer pH 8.0, 1 year, 20% loss of activity Clostridium kluyveri
1.3.8.1 -18°C, 30% ethylene glycol, 15 mM phosphate pH 8.0, 20% loss of activity within 1 year Clostridium kluyveri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.95 (E)-2-butenoate + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 (E)-2-butenoyl-CoA + methyl viologen second best substrate, 45% activity compared to acryloyl-CoA Clostridium kluyveri ?
-
?
1.3.1.95 1-penten-3-one + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 2-cyclohexen-1-one + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 2-methylacrolein + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 4-phenyl-3-buten-2-one + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 acrolein + methyl viologen
-
Clostridium kluyveri ?
-
?
1.3.1.95 acrylic acid + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 acryloyl-CoA + methyl viologen best substrate Clostridium kluyveri ?
-
?
1.3.1.95 butyryl-CoA + 2,6-dichlorophenolindophenol about 1% of the reduction rate of (E)-2-butenoyl-CoA and methyl viologen Clostridium kluyveri ?
-
?
1.3.1.95 diethyl ketone + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 ethyl acrylate + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 ethyl vinyl ketone + methyl viologen
-
Clostridium kluyveri ?
-
?
1.3.1.95 methyl propenyl ketone + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 methyl vinyl ketone + methyl viologen less than 2% activity of that with ethyl vinyl ketone Clostridium kluyveri ?
-
?
1.3.1.95 additional information the enzyme shows no activity with different concentrations of NAD(P)H in 0.1 M phosphate buffer pH 7.0 and ethyl vinyl ketone as substrate Clostridium kluyveri ?
-
?
1.3.1.95 N-acetyl-S-acryloylcysteamine + methyl viologen
-
Clostridium kluyveri ?
-
?
1.3.8.1 (E)-2-butenoyl-CoA + reduced methylviologen
-
Clostridium kluyveri ? + oxidized methylviologen
-
?
1.3.8.1 acrolein + reduced methylviologen
-
Clostridium kluyveri ? + oxidized methylviologen
-
?
1.3.8.1 acryloyl-CoA + reduced methylviologen
-
Clostridium kluyveri propanoyl-CoA + oxidized methylviologen
-
?
1.3.8.1 ethyl vinyl ketone + reduced methylviologen
-
Clostridium kluyveri ? + oxidized methylviologen
-
?
1.3.8.1 N-acetyl-S-acryloyl-cysteamine + reduced methylviologen
-
Clostridium kluyveri ? + oxidized methylviologen
-
?

Subunits

EC Number Subunits Comment Organism
1.3.1.95 homodimer 2 * 14200, SDS-PAGE Clostridium kluyveri
1.3.8.1 dimer 2 * 14200, SDS-PAGE Clostridium kluyveri

Synonyms

EC Number Synonyms Comment Organism
1.3.1.95 acryloyl-CoA reductase
-
Clostridium kluyveri

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.1.95 5.9
-
two optima at pH 5.9 and 6.4 Clostridium kluyveri
1.3.1.95 6.4
-
two optima at pH 5.9 and 6.4 Clostridium kluyveri
1.3.8.1 5.9
-
and 6.4 Clostridium kluyveri
1.3.8.1 6.4
-
and 5.9 Clostridium kluyveri

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.3.1.95 3.9 8.3 the activity of the acryloyl-CoA reductase decreases to about 30% at pH 3.9 or pH 8.3, respectively Clostridium kluyveri
1.3.8.1 3.9
-
30% resiudal activity Clostridium kluyveri
1.3.8.1 8.3
-
30% residual activity Clostridium kluyveri

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.3.8.1 additional information
-
enzyme is more stable at pH 8.0 than at pH 6.0 Clostridium kluyveri

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.95 FMN contains 1.5 mol FMN Clostridium kluyveri
1.3.1.95 additional information he activity of the acryloyl-CoA reductase is not altered significantly by any of the following substances added in concentrations of 0.075 to 0.15 mM: FMN, FAD, AMP, ADP, ATP, NAD+, NADH, acetyl phosphate and co-enzyme A Clostridium kluyveri
1.3.8.1 FMN about 1.5 per mol of dimer Clostridium kluyveri

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.3.1.95 Clostridium kluyveri isoelectric focusing
-
7.5
1.3.8.1 Clostridium kluyveri polyacrylamide slab gel
-
6.9
1.3.8.1 Clostridium kluyveri chromatofocusing
-
7.5