BRENDA - Enzyme Database show
show all sequences of 1.3.1.86

Purification of crotonyl-CoA reductase from Streptomyces collinus and cloning, sequencing and expression of the corresponding gene in Escherichia coli

Wallace, K.K.; Bao, Z.Y.; Dai, H.; Digate, R.; Schuler, G.; Speedie, M.K.; Reynolds, K.A.; Eur. J. Biochem. 233, 954-962 (1995)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
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)
Commentary
Organism
expressed in Escherichia coli BL21 (DE3)/pZYB3 cells
Streptomyces collinus
Inhibitors
Inhibitors
Commentary
Organism
Structure
ammonium sulfate
enzyme activity is inhibited by ammonium sulfatehowever, 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
Commentary
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 30C
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 30C
Streptomyces collinus
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
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
Primary Accession No. (UniProt)
Commentary
Textmining
Streptomyces collinus
-
-
-
Purification (Commentary)
Commentary
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 [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
0.00091
-
native enzyme from crude extract, pH 7.5 at 30C
Streptomyces collinus
0.333
-
recombinant enzyme from crude extract, pH 7.5 at 30C
Streptomyces collinus
2.889
-
native enzyme after 3068fold purification, pH 7.5 at 30C
Streptomyces collinus
3.316
-
recombinant enzyme after 10fold purification, pH 7.5 at 30C
Streptomyces collinus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
crotonyl-CoA + NADPH + H+
the enzyme exhibits a high substrate specificity for crotonyl-CoA
711884
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
711884
Streptomyces collinus
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
homodimer
2 * 48000, SDS-PAGE; 2 * 49400, calculated from amino acid sequence
Streptomyces collinus
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
40
-
-
Streptomyces collinus
Temperature Stability [C]
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
40
-
at 40C the enzyme retains 47% of its activity after 30 min
Streptomyces collinus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
-
Streptomyces collinus
Cofactor
Cofactor
Commentary
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
Commentary
Organism
Structure
0.0095
-
palmitoyl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
0.017
-
myristoyl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
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 30C
Streptomyces collinus
0.5
-
isomyristoyl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
0.63
-
NADP+
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
0.9
-
butyryl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
Activating Compound (protein specific)
Activating Compound
Commentary
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) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21 (DE3)/pZYB3 cells
Streptomyces collinus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
NADPH is the sole electron donor for the reduction catalyzed by crotonyl-CoA reductase
Streptomyces collinus
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
ammonium sulfate
enzyme activity is inhibited by ammonium sulfatehowever, 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
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0095
-
palmitoyl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
0.017
-
myristoyl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
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 30C
Streptomyces collinus
0.5
-
isomyristoyl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
0.63
-
NADP+
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
0.9
-
butyryl-CoA
in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30C
Streptomyces collinus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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 30C
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 30C
Streptomyces collinus
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
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
Purification (Commentary) (protein specific)
Commentary
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] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
0.00091
-
native enzyme from crude extract, pH 7.5 at 30C
Streptomyces collinus
0.333
-
recombinant enzyme from crude extract, pH 7.5 at 30C
Streptomyces collinus
2.889
-
native enzyme after 3068fold purification, pH 7.5 at 30C
Streptomyces collinus
3.316
-
recombinant enzyme after 10fold purification, pH 7.5 at 30C
Streptomyces collinus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
crotonyl-CoA + NADPH + H+
the enzyme exhibits a high substrate specificity for crotonyl-CoA
711884
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
711884
Streptomyces collinus
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 48000, SDS-PAGE; 2 * 49400, calculated from amino acid sequence
Streptomyces collinus
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
40
-
-
Streptomyces collinus
Temperature Stability [C] (protein specific)
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
40
-
at 40C the enzyme retains 47% of its activity after 30 min
Streptomyces collinus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
-
Streptomyces collinus
General Information
General Information
Commentary
Organism
physiological function
the enzyme plays a role in providing butyryl-CoA as a starter unit for straight-chain fatty acid biosynthesis
Streptomyces collinus
General Information (protein specific)
General Information
Commentary
Organism
physiological function
the enzyme plays a role in providing butyryl-CoA as a starter unit for straight-chain fatty acid biosynthesis
Streptomyces collinus
Other publictions for EC 1.3.1.86
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
725287
Aboulnaga
Effect of an Oxygen-Tolerant B ...
Clostridioides difficile
J. Bacteriol.
195
3704-3713
2013
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712235
Liu
Biosynthesis of salinosporamid ...
Salinispora tropica
J. Am. Chem. Soc.
131
10376-10377
2009
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712641
Akopiants
Multiple pathways for acetate ...
Streptomyces cinnamonensis
J. Ind. Microbiol. Biotechnol.
33
141-150
2006
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656737
Li
Crotonyl-coenzyme A reductase ...
Streptomyces cinnamonensis, Streptomyces collinus
Microbiology
150
3463-3472
2004
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1
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711366
Fukui
Engineering of Ralstonia eutro ...
Streptomyces cinnamonensis
Biomacromolecules
3
618-624
2002
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712939
Liu
Precursor supply for polyketid ...
Streptomyces cinnamonensis
Metab. Eng.
3
40-48
2001
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712617
Sun
-
Crotonic acid-directed biosynt ...
Streptomyces hygroscopicus subsp. ascomyceticus
J. Ferment. Bioeng.
86
261-265
1998
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713435
Stassi
Ethyl-substituted erythromycin ...
Streptomyces collinus
Proc. Natl. Acad. Sci. USA
95
7305-7309
1998
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712233
Liu
-
Linking diversity in evolution ...
Streptomyces collinus
J. Am. Chem. Soc.
119
2973-2979
1997
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712270
Han
A novel alternate anaplerotic ...
Streptomyces collinus
J. Bacteriol.
179
5157-5164
1997
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711884
Wallace
Purification of crotonyl-CoA r ...
Streptomyces collinus
Eur. J. Biochem.
233
954-962
1995
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711170
Dodds
-
Effects of coenzyme A and pH o ...
Bos taurus
Biochem. Soc. Trans.
9
556-557
1981
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1
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390850
Strom
-
Activation and inhibition of c ...
Bos taurus
J. Biol. Chem.
254
8159-8162
1979
1
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9
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1
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1
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4
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1
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9
4
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1
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390852
Maitra
Crotonyl coenzyme A reductase ...
Bos taurus
J. Biol. Chem.
249
111-117
1974
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