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Literature summary for 1.3.1.86 extracted from

  • Wallace, K.K.; Bao, Z.Y.; Dai, H.; Digate, R.; Schuler, G.; Speedie, M.K.; Reynolds, K.A.
    Purification of crotonyl-CoA reductase from Streptomyces collinus and cloning, sequencing and expression of the corresponding gene in Escherichia coli (1995), Eur. J. Biochem., 233, 954-962.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information no significant activation of the activity of crotonyl-CoA reductase is observed upon addition of either flavin adenine dinucleotide (0.018 and 0.072 mM) or flavin mononucleotide (0.013 and 0.130 mM) Streptomyces collinus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 (DE3)/pZYB3 cells Streptomyces collinus

Inhibitors

Inhibitors Comment Organism Structure
ammonium sulfate enzyme activity is inhibited by ammonium sulfate ; however, this inhibition is overcome by addition of 10 mM guanidine Streptomyces collinus
arachidoyl-CoA 86% residual activity at 0.1 mM Streptomyces collinus
butyryl-CoA slight inhibition Streptomyces collinus
Ca2+ complete inhibition at 1 mM Streptomyces collinus
Co2+ complete inhibition at 1 mM Streptomyces collinus
iodoacetamide 40% inhibition at 1 mM Streptomyces collinus
isomyristoyl-CoA 78% residual activity at 0.1 mM Streptomyces collinus
isopalmitoyl-CoA 95% residual activity at 0.1 mM Streptomyces collinus
Mg2+ 30% inhibition at 1 mM Streptomyces collinus
Mn2+ complete inhibition at 1 mM Streptomyces collinus
additional information no significant inhibition of the activity of crotonyl-CoA reductase is observed upon addition of either flavin adenine dinucleotide (0.018 and 0.072 mM) or flavin mononucleotide (0.013 and 0.130 mM) Streptomyces collinus
myristoyl-CoA 36% residual activity at 0.1 mM Streptomyces collinus
N-ethylmaleimide 80% inhibition at 1 mM Streptomyces collinus
NADP+
-
Streptomyces collinus
NADPH concentrations of NADPH above 0.2 mM lead to inhibition of enzyme activity Streptomyces collinus
p-chloromercuribenzoate a 30-min incubation of crotonyl-CoA reductase with p-chloromercuribenzoate at 0.008 mM leads to approximately 8.5% inhibition of enzyme activity Streptomyces collinus
palmitoyl-CoA 24% residual activity at 0.1 mM Streptomyces collinus
stearoyl-CoA 92% residual activity at 0.1 mM Streptomyces collinus
Zn2+ 55% inhibition at 1 mM Streptomyces collinus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.015
-
NADPH in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 1 mM dithioerythritol and 10% (v/v) glycerol, at 30°C Streptomyces collinus
0.018
-
crotonyl-CoA in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 1 mM dithioerythritol and 10% (v/v) glycerol, at 30°C Streptomyces collinus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
2 * 48000, SDS-PAGE Streptomyces collinus
49400
-
2 * 49400, calculated from amino acid sequence Streptomyces collinus
85000
-
native protein, gel filtration Streptomyces collinus

Organism

Organism UniProt Comment Textmining
Streptomyces collinus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, DEAE-cellulose column chromatography, phenyl-Sepharose column chromatography, Mono Q column chromatography, Sephadex G-100 gel filtration, and phenyl-Superose gel filtration Streptomyces collinus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.00091
-
native enzyme from crude extract, pH 7.5 at 30°C Streptomyces collinus
0.333
-
recombinant enzyme from crude extract, pH 7.5 at 30°C Streptomyces collinus
2.889
-
native enzyme after 3068fold purification, pH 7.5 at 30°C Streptomyces collinus
3.316
-
recombinant enzyme after 10fold purification, pH 7.5 at 30°C Streptomyces collinus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
crotonyl-CoA + NADPH + H+ the enzyme exhibits a high substrate specificity for crotonyl-CoA Streptomyces collinus butanoyl-CoA + NADP+
-
?
additional information the enzyme is unable to catalyze the reduction of any other enoyl-CoA thioesters (acryloyl-CoA, trans-2-pentenoyl-CoA, trans-hexenoyl-CoA, trans-2-octenoyl-CoA, trans-2-dodecenoyl-CoA, trans-2-hexadecenoyl-CoA) or to utilize NADH as an electron donor. The enzyme is unable to reduce either the N-acetylcysteamine or the pantetheine thioester of crotonic acid Streptomyces collinus ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 48000, SDS-PAGE Streptomyces collinus
homodimer 2 * 49400, calculated from amino acid sequence Streptomyces collinus

Synonyms

Synonyms Comment Organism
acyl-CoA:NADP+ trans-2-oxidoreductase
-
Streptomyces collinus
crotonyl-CoA reductase
-
Streptomyces collinus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Streptomyces collinus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
at 40°C the enzyme retains 47% of its activity after 30 min Streptomyces collinus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Streptomyces collinus

Cofactor

Cofactor Comment Organism Structure
NADPH NADPH is the sole electron donor for the reduction catalyzed by crotonyl-CoA reductase Streptomyces collinus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0095
-
palmitoyl-CoA in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C Streptomyces collinus
0.017
-
myristoyl-CoA in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C Streptomyces collinus
0.4
-
isopalmitoyl-CoA Ki above 0.4 mM, in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C Streptomyces collinus
0.5
-
isomyristoyl-CoA in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C Streptomyces collinus
0.63
-
NADP+ in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C Streptomyces collinus
0.9
-
butyryl-CoA in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C Streptomyces collinus

General Information

General Information Comment Organism
physiological function the enzyme plays a role in providing butyryl-CoA as a starter unit for straight-chain fatty acid biosynthesis Streptomyces collinus