Information on EC 1.1.1.35 - 3-hydroxyacyl-CoA dehydrogenase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY hide
1.1.1.35
-
RECOMMENDED NAME
GeneOntology No.
3-hydroxyacyl-CoA dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(S)-3-hydroxyacyl-CoA + NAD+ = 3-oxoacyl-CoA + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
(R)- and (S)-3-hydroxybutanoate biosynthesis (engineered)
-
-
2-methylpropene degradation
-
-
3-hydroxypropanoate/4-hydroxybutanate cycle
-
-
4-hydroxybenzoate biosynthesis V
-
-
androstenedione degradation
-
-
benzoyl-CoA degradation I (aerobic)
-
-
cholesterol degradation to androstenedione I (cholesterol oxidase)
-
-
cholesterol degradation to androstenedione II (cholesterol dehydrogenase)
-
-
crotonate fermentation (to acetate and cyclohexane carboxylate)
-
-
fatty acid beta-oxidation I
-
-
fatty acid beta-oxidation II (peroxisome)
-
-
fatty acid beta-oxidation VI (peroxisome)
-
-
fatty acid salvage
-
-
glutaryl-CoA degradation
-
-
jasmonic acid biosynthesis
-
-
L-glutamate degradation V (via hydroxyglutarate)
-
-
methyl ketone biosynthesis (engineered)
-
-
methyl tert-butyl ether degradation
-
-
phenylacetate degradation I (aerobic)
-
-
pyruvate fermentation to butanoate
-
-
pyruvate fermentation to butanol I
-
-
pyruvate fermentation to butanol II (engineered)
-
-
pyruvate fermentation to hexanol (engineered)
-
-
toluene degradation to benzoyl-CoA (anaerobic)
-
-
adipate degradation
-
-
butanoate fermentation
-
-
CO2 fixation in Crenarchaeota
-
-
lipid metabolism
-
-
phenylacetate degradation (aerobic)
-
-
tryptophan metabolism
-
-
Fatty acid elongation
-
-
Fatty acid degradation
-
-
Primary bile acid biosynthesis
-
-
Valine, leucine and isoleucine degradation
-
-
Geraniol degradation
-
-
Lysine degradation
-
-
Benzoate degradation
-
-
Tryptophan metabolism
-
-
Toluene degradation
-
-
Butanoate metabolism
-
-
Carbon fixation pathways in prokaryotes
-
-
Caprolactam degradation
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
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-
Microbial metabolism in diverse environments
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-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
(S)-3-hydroxyacyl-CoA:NAD+ oxidoreductase
Also oxidizes S-3-hydroxyacyl-N-acylthioethanolamine and S-3-hydroxyacyl-hydrolipoate. Some enzymes act, more slowly, with NADP+. Broad specificity to acyl chain-length (cf. EC 1.1.1.211 [long-chain-3-hydroxyacyl-CoA dehydrogenase]).
CAS REGISTRY NUMBER
COMMENTARY hide
9028-40-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
multifunctional beta-oxidation enzyme complex composed of 2 different 3-hydroyacyl-CoA dehydrogenases, 2 different 2-enoyl-CoA hydratases, thiolase, and epimerase activities
-
-
Manually annotated by BRENDA team
multifunctional beta-oxidation enzyme complex composed of 2 different 3-hydroyacyl-CoA dehydrogenases, 2 different 2-enoyl-CoA hydratases, thiolase, and epimerase activities
-
-
Manually annotated by BRENDA team
gene FUM13
-
-
Manually annotated by BRENDA team
gene FUM13
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene fadB
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(3S)-3-hydroxyadipyl-CoA + NAD+
3-oxoadipyl-CoA + NADH + H+
show the reaction diagram
(R)-3-hydroxyacyl-CoA + NAD+
3-oxoacyl-CoA + NADH + H+
show the reaction diagram
-
cf. EC 1.1.1.36
-
-
r
(S)-3-hydroxyacyl-CoA + NAD+
3-oxoacyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxybutanoyl-CoA + NAD+
acetoacetyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxybutanoyl-CoA + NADP+
acetoacetyl-CoA + NADPH + H+
show the reaction diagram
(S)-3-hydroxybutyryl-CoA + NAD+
3-acetoacetyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxybutyryl-CoA + NAD+
3-oxobutyryl-CoA + NADH + H+
show the reaction diagram
-
-
-
-
?
(S)-3-hydroxybutyryl-CoA + NAD+
acetoacetyl-CoA + NADH
show the reaction diagram
(S)-3-hydroxybutyryl-CoA + NAD+
acetoacetyl-CoA + NADH + H+
show the reaction diagram
-
-
-
-
?
(S)-3-hydroxybutyryl-CoA + NADP+
acetoacetyl-CoA + NADPH
show the reaction diagram
(S)-3-hydroxydecanoyl-CoA + ?
3-oxodecanoyl-CoA + NADH + H+
show the reaction diagram
-
-
-
-
?
(S)-3-hydroxydecanoyl-CoA + NAD+
3-ketodecanoyl-CoA + NADH
show the reaction diagram
(S)-3-hydroxyhexanoyl-CoA + NAD+
3-ketohexanoyl-CoA + NADH
show the reaction diagram
(S)-3-hydroxyhexenoyl-CoA + NAD+
3-ketohexenoyl-CoA + NADH
show the reaction diagram
-
-
-
r
(S)-3-hydroxylauryl-CoA + NAD+
3-ketolauryl-CoA + NADH
show the reaction diagram
-
-
-
r
1-propanol + NAD+
n-propanal + NADH
show the reaction diagram
-
multifunctional enzyme from brain
-
ir
17beta-estradiol + NAD+
estrone + NADH
show the reaction diagram
17beta-estradiol + NAD+
estrone + NADH + H+
show the reaction diagram
2-fluoro-3-hydroxy-4-octenoyl-CoA + NAD+
2-fluoro-3-oxo-4-octenoyl-CoA + NADH
show the reaction diagram
-
-
-
-
r
2-fluoro-3-hydroxyoctanoyl-CoA + NAD+
2-fluoro-3-oxooctanoyl-CoA + NADH
show the reaction diagram
-
-
-
-
r
2-propanol + NAD+
acetone + NADH
show the reaction diagram
-
multifunctional enzyme from brain
-
ir
3-acetoacetyl-CoA + NADH + H+
(S)-3-hydroxybutyryl-CoA + NAD+
show the reaction diagram
3-acetoacetyl-CoA + NADPH + H+
(S)-3-hydroxybutyryl-CoA + NAD+
show the reaction diagram
3-hydroxy-2-methylacyl-CoA + NAD+
3-oxo-2-methylacyl-CoA + NADH
show the reaction diagram
-
preferred substrate of SCHAD, no activity towards 3-hydroxy-2-methylacyl-CoA by HAD
-
-
-
3-hydroxy-4-octenoyl-CoA + NAD+
3-oxo-4-octenoyl-CoA + NADH
show the reaction diagram
-
-
-
-
r
3-hydroxybutyryl-CoA + NAD+
acetoacetyl-CoA + NADH
show the reaction diagram
3-hydroxyoctanoyl-CoA + NAD+
3-oxooctanoyl-CoA + NADH
show the reaction diagram
-
-
-
-
r
3-ketohexadecanoyl-CoA + NADH
(S)-3-hydroxhexadecanoyl-CoA + NAD+
show the reaction diagram
3-ketooctanoyl-CoA + NADH
(S)-3-hydroxyoctanoyl-CoA + NAD+
show the reaction diagram
3-oxoacyl-CoA + NADH
3-hydroxyacyl-CoA + NAD+
show the reaction diagram
-
key enzyme involved in fatty acid oxidation
-
-
r
3-oxofumonisin B3 + NADPH
fumonisin B3 + NADP+
show the reaction diagram
3-oxohexadecanoyl-CoA + NADH + H+
(S)-3-hydroxhexadecanoyl-CoA + NAD+
show the reaction diagram
-
-
-
-
?
3-oxooctanoyl-CoA + NADH + H+
(S)-3-hydroxyoctanoyl-CoA + NAD+
show the reaction diagram
-
-
-
-
?
5alpha-androstane-3,17-diol + NAD+
5alpha-dihydrotestosterone + NADH
show the reaction diagram
5alpha-dihydrotestosterone + NADH
(3beta,5alpha,17beta)-androstane-3,17-diol + NAD+
show the reaction diagram
-
-
-
r
8-(acetoacetylthio)-6-ethyloctanoic acid + NADH
6-ethyl-8-[[(1S)-1-hydroxy-3-oxobutyl]thio]octanoic acid + NAD+
show the reaction diagram
-
-
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r
8-(acetoacetylthio)-6-mercaptooctanoic acid + NADH
8-[[(1S)-1-hydroxy-3-oxobutyl]thio]-6-mercaptooctanoic acid + NAD+
show the reaction diagram
-
-
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r
acetoacetyl-CoA + NADH
(S)-3-hydroxybutyryl-CoA + NAD+
show the reaction diagram
acetoacetyl-CoA + NADH
3-hydroxybutyryl-CoA + NAD+
show the reaction diagram
acetoacetyl-CoA + NADH + H+
3-hydroxybutyryl-CoA + NAD+
show the reaction diagram
-
-
-
-
?
acetoacetyl-cysteamine-2,2,5,5-tetramethyl-1-oxy-3-pyrroline-3-carboxylic acid amide + NADH
(S)-3-hydroxybutyryl-cysteamine-2,2,5,5-tetramethyl-1-oxy-3-pyrroline-3-carboxylic acid amide + NAD+
show the reaction diagram
-
-
-
r
acetoacetyl-N-acetylcysteamine + NADH
(S)-3-hydroxybutyryl-N-acetylcysteamine + NAD+
show the reaction diagram
acetoacetyl-N-beta-alanylcysteamine + NADH
(S)-3-hydroxybutyryl-N-beta-alanylcysteamine + NAD+
show the reaction diagram
-
-
-
r
acetoacetyl-pantetheine + NADH
(S)-3-hydroxybutyryl-pantetheine
show the reaction diagram
acetoacetyl-pantetheine-4'-(2,2,5,5-tetramethyl-1-oxy-3-pyrroline-3-carboxylic acid ester) + NADH
(S)-3-hydroxybutyryl-pantetheine-4'-(2,2,5,5-tetramethyl-1-oxy-3-pyrroline-3-carboxylic acid ester) + NAD+
show the reaction diagram
-
-
-
r
acetoacetyldecanoate + NADH
(S)-3-hydroxybutyryldecanoate + NAD+
show the reaction diagram
-
-
-
r
allopregnanolone + NAD+
5alpha-dihydroprogesterone + NADH
show the reaction diagram
androsterone + NAD+
androstanedione + NADH
show the reaction diagram
-
-
-
ir
tiglyl-CoA + NAD+
3-oxo-2-methylacyl-CoA + NADH
show the reaction diagram
-
activity of SCHAD
-
-
-
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(3S)-3-hydroxyadipyl-CoA + NAD+
3-oxoadipyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxyacyl-CoA + NAD+
3-oxoacyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxybutanoyl-CoA + NAD+
acetoacetyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxybutyryl-CoA + NAD+
3-acetoacetyl-CoA + NADH + H+
show the reaction diagram
(S)-3-hydroxybutyryl-CoA + NAD+
acetoacetyl-CoA + NADH
show the reaction diagram
17beta-estradiol + NAD+
estrone + NADH
show the reaction diagram
-
enzyme conatains 17beta-hydroxysteroid and 3alpha-hydroxysteroid dehydrogenase activity
-
ir
17beta-estradiol + NAD+
estrone + NADH + H+
show the reaction diagram
-
inactivation
-
-
?
3-hydroxybutyryl-CoA + NAD+
acetoacetyl-CoA + NADH
show the reaction diagram
3-oxoacyl-CoA + NADH
3-hydroxyacyl-CoA + NAD+
show the reaction diagram
-
key enzyme involved in fatty acid oxidation
-
-
r
3-oxofumonisin B3 + NADPH
fumonisin B3 + NADP+
show the reaction diagram
5alpha-androstane-3,17-diol + NAD+
5alpha-dihydrotestosterone + NADH
show the reaction diagram
-
inactivation
-
-
?
allopregnanolone + NAD+
5alpha-dihydroprogesterone + NADH
show the reaction diagram
-
inactivation
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
NADPH
additional information
the enzyme prefers NAD+. NADP(H) is utilized at a rate of less than 10% in comparison to activity with NAD(H)
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
enhances activity at 1 mM or 10 mM
Mg2+
enhances activity at 1 mM or 10 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
acetyl-CoA
inhibits the enzyme by 50% at 0.1 mM, further increase of inhibitor concentration reduces the activity of FadB' drastically
Co2+
1 mM, about 50% inhibition
CoA
inhibits the enzyme by 20% at 0.1 mM, further increase of inhibitor concentration reduces the activity of FadB' drastically
EDTA
about 35% inhibition at 1 mM, about 25% inhibition at 10 mM
iodoacetamide
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-
iodoacetic acid
-
-
K+
about 30% inhibition at 1 mM, about 20% inhibition at 10 mM
N-bromsuccinimide
-
-
N-ethylmaleimide
-
-
Ni2+
1 mM, about 60% inhibition
p-chloromercuribenzoate
-
-
p-Chloromercuriphenyl sulfonic acid
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95% inhibition at 0.1 mM
RNAi
-
suppressed KCR activity results in a reduction of cuticular wax load and affects very-long-chain fatty acid composition of sphingolipids, seed triacylglycerols, and root glycerolipids
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Zn2+
1 mM, about 60% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
AMP-activated protein kinase
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-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.034
(3beta,5alpha,17beta)-androstane-3,17-diol
-
enzyme from brain
0.0505 - 0.1157
(R)-3-hydroxyacyl-CoA
0.06 - 0.2
(S)-3-hydroxybutanoyl-CoA
0.000058 - 43.5
(S)-3-hydroxybutyryl-CoA
0.088
(S)-3-hydroxydecanoyl-CoA
-
mitochondrial enzyme, oxidation of (S)-3-hydroxydecanoyl-CoA
0.0286 - 0.34
(S)-3-hydroxyhexanoyl-CoA
0.1
(S)-3-hydroxylauryl-CoA
-
-
0.0163 - 0.35
(S)-3-hydroxyoctanoyl-CoA
0.043
17beta-estradiol
-
enzyme from brain
65.6
3-acetoacetyl-CoA
pH 7.0, 30C
0.003 - 0.263
acetoacetyl-CoA
44.4
acetoacetyl-N-acetyl-cysteamine
-
-
10
acetoacetyl-N-acetylcysteamine
-
-
1.4
acetoacetyl-N-beta-alanylcysteamine
-
-
10
acetoacetyl-Nacetylcysteamine
-
-
0.08 - 1.19
acetoacetyl-pantetheine
0.045
androsterone
-
enzyme from brain
0.00009 - 29.5
NAD+
0.0009 - 50
NADH
11
NADP+
-
-
0.435
NADPH
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.093
(3beta,5alpha,17beta)-androstane-3,17-diol
-
enzyme from brain
2 - 45
(R)-3-hydroxyacyl-CoA
15
(S)-3-hydroxybutanoyl-CoA
-
pH 8.0, 70C
0.012 - 1420
(S)-3-hydroxybutyryl-CoA
0.011
17beta-estradiol
-
enzyme from brain
0.011
3-acetoacetyl-CoA
pH 7.0, 30C
45
3-ketohexadecanoyl-CoA
-
-
24
3-ketohexdecanoyl-CoA
enzyme from brain, pH 7.0
28 - 102
3-ketooctanoyl-CoA
0.019 - 45500
acetoacetyl-CoA
0.011
androsterone
-
enzyme from brain
0.004 - 1817
NAD+
0.189 - 510
NADH
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
90 - 600
(R)-3-hydroxyacyl-CoA
260
(S)-3-hydroxybutanoyl-CoA
-
pH 8.0, 70C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.002
-
enzyme from brain, substrate 1-propanol
0.0121
-
enzyme from brain, substrate androsterone
0.0156
-
enzyme from brain, substrate 17beta-estradiol
0.033
-
enzyme activity in brain mitochondria, substrate acetoacetyl-CoA
0.087
-
enzyme from brain, substrate 5alpha-dihydrotestosterone
0.13
-
enzyme from brain, substrate dihydroandrosterone
0.41
-
enzyme activity in solubilized sediments of homogenized cells
0.48
-
enzyme activity in inner membrane of liver mitochondria, substrate acetoacetyl-CoA
0.5
85C, pH 7.0, highly enriched enzyme
0.75
-
72% activivty is bound to the matrix surface of the inner mitochondria membrane, binding is inhibited by increasing ionic strength and pH
0.78
-
enzyme activity in kidney mitochondria, substrate acetoacetyl-CoA
1.67
-
enzyme activity in liver mitochondria, substrate acetoacetyl-CoA
2.3
-
enzyme activity in liver mitoplasts, substrate acetoacetyl-CoA
2.77
-
trans-3-decenoyl-CoA as a substrate
3 - 8
-
pH 8.0, 65C, native enzyme, autotrophically- or heterotrophically-grown cell
3.4
-
enzyme activity in matrix of liver mitochondria, substrate acetoacetyl-CoA
4.17
-
(S)-3-hydroxydecanoyl-CoA as substrate, activity of L-specific 3-hydroxyacyl-CoA dehydrogenase in perMFE-I
4.22
-
(S)-3-hydroxybutyryl-CoA as substrate, activity of L-specific 3-hydroxyacyl-CoA dehydrogenase in perMFE-I
12.6
-
peroxisomal enzyme
16.5
purified recombinant enzyme, pH 6.5, 30C, substrate (S)-3-hydroxybutanoyl-CoA with NAD+
17.35
enzyme from brain
23.4
-
L-3-hydroxyacyl-CoA dehydrogenase activity of the trifunctional beta-oxidation protein
40
-
purified enzyme
176
-
-
220
-
-
452
-
purified recombinant His-tagged wild-type enzyme
511
-
recombinant enzyme
565
-
mitochondrial enzyme
1200
-
His-tagged recombinant enzyme
additional information
-
the N-terminal His-tag does notinfluence enzyme activity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
mitochondrial enzyme, reduction of acetoacetyl-CoA
5
-
phosphate buffer
5.5 - 6.5
conversion of acetoacetyl-CoA to beta-hydroxybutyryl-CoA decreases above pH 6.5
6
-
reduction of acetoacetyl-CoA
6 - 7
-
reduction of acetoacetyl-CoA
6.2
-
peroxisomal enzyme, reduction of acetoacetyl-CoA
6.5 - 7
-
enzyme from brain, reduction of acetoacetyl-CoA
7.2
assay at; assay at
8.5
-
SCAD I, pH optimum for oxidation
9
-
oxidation of (S)-3-hydroxybutyryl-CoA
9.3
-
SCHAD II, pH optimum for oxidation
9.5 - 10
-
enzyme from brain, oxidation of 17beta-estradiol
9.8
-
peroxisomal enzyme, oxidation of (S)-3-hydroxybutyryl-CoA
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 10
maximal activity at pH 6.0-7.0, 65% and 75% activity at pH 5.0 and pH 8.0, respectively, and 46% of the maximal activity at pH 10.0, and 26% of maximal activity at pH 4.0
9 - 11
pH 9.0: about 45% of maximal activity, pH 11.0: about 55% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
calculated, native protein
6.1
calculated, recombinant protein
7.66
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
activated, high level expression
Manually annotated by BRENDA team
-
KCR1 and KCR2 transcripts
Manually annotated by BRENDA team
-
KCR1 and KCR2 transcripts
Manually annotated by BRENDA team
-
activity of the enzyme it encodes is particularly high in the pancreas and especially in the islets of Langerhans
Manually annotated by BRENDA team
-
KCR1 and KCR2 transcripts
Manually annotated by BRENDA team
-
in oocytes from diabetic mice, activity of Hadh2 is significantly reduced
Manually annotated by BRENDA team
-
high level expression in malignant prostatic epithelial cells
Manually annotated by BRENDA team
-
KCR1 and KCR2 transcripts
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Cupriavidus necator (strain ATCC 17699 / H16 / DSM 428 / Stanier 337)
Q0KEY8
Cupriavidus necator (strain ATCC 17699 / H16 / DSM 428 / Stanier 337)
Q0KEY8
Cupriavidus necator (strain ATCC 17699 / H16 / DSM 428 / Stanier 337)
Q0KEY8
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27000
4 * 27000, enzyme from brain, SDS-PAGE
33000
-
2 * 33000, SDS-PAGE
35000
x * 35000, SDS-PAGE; x * 35000, SDS-PAGE
40000
-
liver enzyme
42000
-
2 * 78000 + 2 * 42000, SDS-PAGE
44500
-
1 * 45500 + 1 * 44500 + 1 * 34000 + 1 * 32000, SDS-PAGE
45500
-
1 * 45500 + 1 * 44500 + 1 * 34000 + 1 * 32000, SDS-PAGE
50300
-
gel filtration
66000
-
gel filtration
76000
-
gel filtration
77000
-
1 * 77000, peroxisomal enzyme, SDS-PAGE
78000
-
2 * 78000 + 2 * 42000, SDS-PAGE
87000
recombinant His-tagged enzyme, gel filtration
93000
-
4 * 93000, trifunctional beta-oxidation protein with activities of EC 1.1.1.35, EC 4.2.1.17 and EC 5.1.2.3, SDS-PAGE
108000
enzyme from brain, gel filtration
260000
-
gel filtration
270000
-
gel filtration
365000
-
trifunctional beta-oxidation protein with activities of EC 1.1.1.35, EC 4.2.1.17 and EC 5.1.2.3, gel filtration
460000
-
native enzyme complex, sucrose density gradient centrifugation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant detagged enzyme, hanging drop vapour diffusion method, 10 mg/ml protein in 25 mM sodium acetate trihydrate, pH 5.0, and 300 mM sodium chloride is mixed with an equal volume of reservoir solution, containing 21% PEG 3350, 0.2 M sodium chloride, 0.1 M MES, pH 6.5, for parallelepiped-shaped crystals and 23% PEG 3350, 0.2 M sodium chloride, 0.1 M bicine, pH 8.0, for cuboid-shaped crystals, equilibration against 0.20 ml reservoir solution, 3-5days at 18C, X-ray diffraction structure determination and analysis of parallelepiped-shaped cuboid-shaped crystal at 1.60 A and 2.20 A resolution, respectively
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purified recombinant detagged wild-type enzyme, crystals are grown by hanging drop method from 23% PEG 3350, 0.2 M sodium chloride, 0.1M N,N-bis (2-hydroxyethyl)glycine, pH 8.0, X-ray diffraction structure determination and analysis at 2.20 A resolution, molecular replacement with the human HAD structure as search model, PDB ID 3had
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hanging drop vapour diffusion method, mixing of 30 mg/ml protein in 40 mM Tris-HCl, pH 8.0, 1 mM DTT, with reservoir solution containing 2 M ammonium sulfate, 0.1 M CAPS, pH 10.5, and 0.2 M lithium sulfate, 22C, 7 days, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement method and structure modeling
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purified recombinant His6-tagged wild-type enzyme in apoform and in complex with substrates acetoacetyl-CoA and NAD+, hanging drop vapour diffusion method, mixing of 30 mg/ml protein in 40 mM Tris-HCl, pH 8.0, 1 mM DTT, with or without 20 mM NAD+, and 20 mM acetoacetyl-CoA, with reservoir solution containing 0.2 M Li2SO4, 0.1 M CAPS, pH 10.5, and 2 M ammonium sulfate, 22C, 7 days, X-ray diffraction structure determination and analysis at 1.8-2.54 A resolution, molecular replacement and structure modeling
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purified recombinant enzyme in apoform, as selenomethionine-labeled enzyme, and in complex with substrates acetoacetyl-CoA and NAD+, hanging drop vapour diffusion method, mixing of 50 mg/ml wild-type protein or selenomethionine-labeled enzyme in 40 mM Tris-HCl, pH 8.0, 1 mM DTT, with reservoir solution containing 2 M ammonium sulfate, 0.1 M sodium cacodylate, pH 6.5, and 0.2 M sodium chloride, 22C, 7 days, X-ray diffraction structure determination and analysis at 2.42-2.7 A resolution, molecular replacement using the crystal structure of the apo-form of RePaaH1, and structure modeling
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purified recombinant wild-type and selenomethionine-labeled enzymes, hanging drop vapour diffusion method, mixing of 30 mg/ml wild-type protein or 25 mg/ml selenomethionine-labeled enzyme in 40 mM Tris-HCl, pH 8.0, 1 mM DTT, with reservoir solution containing 1.4 M ammonium sulfate, 0.1 M sodium cacodylate, pH 6.0, and 0.2 M sodium chloride, 22C, 7 days, X-ray diffraction structure determination and analysis at 2.6 A resolution, single-wavelength anomalous dispersion method
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50 mM N(2-acetamido)-2-iminodiacetic acid, pH 6.5, polyethylene glycol 4000, 5 mM NAD+ hanging drop, crystals within 3 to 5 days at 18C, enzyme structure is compromised of two domains, a NAD+-binding domain and a helical C-terminal domain
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50% saturation with ammonium sulfate solution, 0.1 M potassium phosphate, pH 6.8, 1 mM EDTA, 2 mM beta-mercaptoethanol, 4C, crystals appear after 2 days
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dialysis against 40% saturated ammonium sulfate containing 100 mM phosphate, 2 mM beta-mercaptoethanol, 1 mM EDTA, pH 6.9, 7.5 or 8.2, vapor diffusion crystallization, crystals are obtained in the ammonium sulfate saturation range of 41% to 48%
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polyethylene glycol, pH 8, orthorhombic crystals, 2.7 A resolution, crystallisation at pH 5 leads to trigonal space group
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two dimers of the enzyme in the asymmetric unit of an orthorombic cell, two coenzyme binding sites per dimer
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 120 mM potassium phosphate, pH 7.0, several months without loss of activity
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-76C, 0.2 M potassium phosphate, pH 6.6, 25% glycerol, 10 mM mercaptoethanol, several months, no loss in activity
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-80C, purified recombinant His-tagged wild-type and mutant enzymes, stable for at least 3 months
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4C, 90% ammonium sulfate, at least 6 monts, no loss of activity
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4C, purified recombinant His-tagged wild-type enzyme, 1 week, no significant loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
60C for 10 min, phosphocellulose, Sephacryl S200
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ammonium sulfate, 50C for 20 min, Zn(OH)2-gel
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ammonium sulfate, first CM-cellulose, gelfiltration, second and third CM-cellulose
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ammonium sulfate, Sephadex G-150, hydroxyapatite, NAD-Sepharose 4B
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beta-oxidation enzyme complex 67fold to homogeneity by ammonium sulfate precipitation, density gradient centrifugation, and ion-exchange chromatography
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calcium phosphate gel, ammonium sulfate, CM-Sephadex, Blue Dextran-Sepharose 4B, calcium phosphate gel-cellulose, Blue Dextran-Sepharose 4B, mitochondrial enzyme; phosphocellulose, ammonium sulfate, CM-cellulose, ammonium sulfate, peroxisomal enzyme
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overexpressed in Escherichia coli, hydroxylapatite
overexpressed in Escherichia coli, phosphocellulose
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recombinant C-terminally GST-tagged enzyme from Escherichia coli strain BL21(DE3) by glutathione affinity and anion exchange chromatography, with cleavage of the GST-tag
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recombinant C-terminally GST-tagged enzyme from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography, cleavage of the GST-tag by PPase, followed by gel filtration, anion exchange chromatography, and ultrafiltration
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recombinant C-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain B834 by nickel affinity chromatography and gel filtration to about 95% purity
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recombinant C-terminally His6-tagged wild-type or selenomethionine-labeled enzymes from Escherichia coli strain B834 by nickel affinity chromatography and gel filtration
Q0KEY8
recombinant His-tagged enzyme from Escherichia coli by Co2+ affinity chromatography
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
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recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography
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recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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recombinant His6-tagged enzyme from Escherichia coli strain B834 by nickel affinity chromatography and gel filtration
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trifunctional beta-oxidation protein with activities of EC 1.1.1.35, EC 4.2.1.17 and EC 5.1.2.3
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
C-terminal hexameric histidine tag, expressed in Escherichia coli
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DNA and amino acid sequence determination and analysis, genetic organization, sequence comparisons and phylogenetic tree of 3-hydroxyacyl-CoA dehydrogenases, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)
DNA and amino acid sequence determination and analysis, recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain B834
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expressed in Escherichia coli
expressed in yeast ybr159DELTA mutant
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expression in Escherichia coli
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
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gene Bn-kcr1, DNA library screening, DNA and amino acid sequence determination and analysis, expression analysis, expression in Saccharomyces cerevisiae BY4742 mutant strains, functional complementation study, overview; gene Bn-kcr2, DNA library screening, DNA and amino acid sequence determination and analysis, expression analysis, expression in Saccharomyces cerevisiae BY4742 mutant strains, functional complementation study, overview
gene F54C8.1, recombinant expression of C-terminally GST-tagged enzyme in Escherichia coli strain BL21(DE3)
gene FUM13, expression of His-tagged enzyme in Escherichia coli, expression in and functional complementation of Saccharomyces cerevisiae 3-ketosphinganine reductase mutant strain tsc10, overview
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gene hadh, quantitative real-time PCR expression analysis
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gene HADHSC, located on chromosome 4q22-26
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gene paaH1, recombinant expression of C-terminally His6-tagged wild-type or selenomethionine-labeled enzymes in Escherichia coli strain B834
Q0KEY8
overexpression in Escherichia coli
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) as soluble N-terminally His-tagged enzymes
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recombinant expression of His-tagged enzyme in Escherichia coli
recombinant expression of His6-tagged enzyme in Escherichia coli strain B834
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recombinantly expressed in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R204A
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site-directed mutagenesis, the mutant with attenuated interactions on the dimerization interface still maintains a dimerization form, but the enzymatic activity is significantly decreased compared the wild-type
R204A/Y209A
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site-directed mutagenesis, the mutant with attenuated interactions on the dimerization interface still maintains a dimerization form, but the enzymatic activity is significantly decreased compared the wild-type
Y209A
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site-directed mutagenesis, the mutant with attenuated interactions on the dimerization interface still maintains a dimerization form, but the enzymatic activity is significantly decreased compared the wild-type
K50A
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site-directed mutagenesis, the mutant shows about 2fold increased activity compared to the wild-type enzyme
K50A//L232Y
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site-directed mutagenesis, the mutant shows about 3fold increased activity compared to the wild-type enzyme
K50A/K54
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site-directed mutagenesis, the mutant shows about 3fold increased activity compared to the wild-type enzyme
K50A/K54A/L232Y
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site-directed mutagenesis, the mutant shows about 5fold increased activity compared to the wild-type enzyme
K54A
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site-directed mutagenesis, the mutant shows about 2fold increased activity compared to the wild-type enzyme
K54A/L232Y
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site-directed mutagenesis, the mutant shows about 4fold increased activity compared to the wild-type enzyme
L232Z
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site-directed mutagenesis, the mutant shows about 2.5fold increased activity compared to the wild-type enzyme
K56A
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site-directed mutagenesis, the mutant shows about twofold increased activity compared to the wild-type enzyme
N190A
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site-directed mutagenesis, the mutant shows highly decreased activity compared to the wild-type enzyme
R52A
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site-directed mutagenesis, the mutant shows highly decreased activity compared to the wild-type enzyme
S119A
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site-directed mutagenesis, almost inactive mutant
K56A
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site-directed mutagenesis, the mutant shows about twofold increased activity compared to the wild-type enzyme
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N190A
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site-directed mutagenesis, the mutant shows highly decreased activity compared to the wild-type enzyme
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R52A
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site-directed mutagenesis, the mutant shows highly decreased activity compared to the wild-type enzyme
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S119A
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site-directed mutagenesis, almost inactive mutant
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D279E
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naturally occuring polymorphism probably involved in development of type 2 diabetes
D45G/Y214H
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a naturally occuring enzyme mutation causing human disease
H152Q