Information on EC 1.3.1.44 - trans-2-enoyl-CoA reductase (NAD+)

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

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EC NUMBER
COMMENTARY hide
1.3.1.44
-
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RECOMMENDED NAME
GeneOntology No.
trans-2-enoyl-CoA reductase (NAD+)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acyl-CoA + NAD+ = trans-didehydroacyl-CoA + NADH + H+
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dehydrogenation
-
-
-
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oxidation
-
-
-
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redox reaction
-
-
-
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reduction
-
-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-leucine degradation IV (Stickland reaction)
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pyruvate fermentation to butanol II (engineered)
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pyruvate fermentation to hexanol (engineered)
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Butanoate metabolism
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Metabolic pathways
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
acyl-CoA:NAD+ trans-2-oxidoreductase
The enzyme from Euglena gracilis acts on crotonoyl-CoA and, more slowly, on trans-hex-2-enoyl-CoA and trans-oct-2-enoyl-CoA.
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CAS REGISTRY NUMBER
COMMENTARY hide
77649-64-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain D1
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Manually annotated by BRENDA team
enzyme is a component of fatty acid synthase type II
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Manually annotated by BRENDA team
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E)-2-hexanoyl-CoA + NADPH
hexanoyl-CoA + NADP+
show the reaction diagram
2-dodecenoyl-CoA + NADH
dodecanoyl-CoA + NAD+
show the reaction diagram
acyl-CoA + NAD+
trans-2,3-didehydroacyl-CoA + NADH
show the reaction diagram
crotonyl-CoA + NAD(P)H
butyryl-CoA + NAD(P)+
show the reaction diagram
-
-
-
?
crotonyl-CoA + NADH
butanoyl-CoA + NAD+
show the reaction diagram
crotonyl-CoA + NADH
butyryl-CoA + NAD+
show the reaction diagram
crotonyl-CoA + NADH + H+
butyryl-CoA + NAD+
show the reaction diagram
crotonyl-CoA + NADPH
butanoyl-CoA + NADP+
show the reaction diagram
crotonyl-CoA + NADPH + H+
butyryl-CoA + NADP+
show the reaction diagram
hexenoyl-CoA + NADH
? + NAD+
show the reaction diagram
trans-2-decenoyl-CoA + NADH
decanoyl-CoA + NAD+
show the reaction diagram
trans-2-dodecenoyl-CoA + NADH
dodecanoyl-CoA + NAD+
show the reaction diagram
trans-2-dodecenoyl-CoA + NADH + H+
dodecanoyl-CoA + NAD+
show the reaction diagram
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-
-
-
?
trans-2-hexadecenoyl-CoA + NADH
hexadecanoyl-CoA + NAD+
show the reaction diagram
trans-2-hexenoyl-CoA + NAD(P)H
hexanoyl-CoA + NAD(P)+
show the reaction diagram
trans-2-hexenoyl-CoA + NADH + H+
hexanoyl-CoA + NAD+
show the reaction diagram
-
-
-
-
?
trans-2-octenoyl-CoA + NADH
octanoyl-CoA + NAD+
show the reaction diagram
trans-didehydroacyl-CoA + NAD(P)H
acyl-CoA + NAD(P)+
show the reaction diagram
i.e. enoyl-CoA
-
-
?
additional information
?
-
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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
acyl-CoA + NAD+
trans-2,3-didehydroacyl-CoA + NADH
show the reaction diagram
crotonyl-CoA + NADH
butanoyl-CoA + NAD+
show the reaction diagram
trans-didehydroacyl-CoA + NAD(P)H
acyl-CoA + NAD(P)+
show the reaction diagram
Q5EU90
i.e. enoyl-CoA
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-
?
additional information
?
-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
flavin
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FAD, riboflavin and FMN increase activity
NADPH
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
butyryl-CoA
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Co2+
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only after preincubation with cation
crotonoyl-CoA
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Hexanoyl-CoA
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isoniazid
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drug to treat tuberculosis
KCN
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protection by FAD
lauroyl-CoA
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-
Mn2+
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only after preincubation with cation
N-ethylmaleimide
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protection by FAD
NAD+
-
competitive to NADH
Ni2+
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only after preincubation with cation
p-chloromercuribenzoate
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protection by FAD
Phenylglyoxal
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-
Zn2+
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only after preincubation with cation
additional information
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2327
(2E)-oct-2-enoyl-CoA
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-
0.0011 - 0.004
2-dodecenoyl-CoA
0.0027
crotonoyl-CoA
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in potassium phosphate buffer pH 6.2, at 30°C
0.049 - 0.21
crotonyl-CoA
0.0893
decenoyl-CoA
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-
0.0558
dodec-2-enoyl-CoA
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-
0.0295
hexadecenoyl-CoA
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-
0.0034 - 0.038
Hexanoyl-CoA
0.0052 - 0.1323
NADH
0.119 - 0.19
NADPH
0.0011 - 0.004
trans-2-dodecenoyl-CoA
0.0034 - 0.091
trans-2-hexenoyl-CoA
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6 - 90
2-dodecenoyl-CoA
0.73 - 92
crotonyl-CoA
15.3 - 112
Hexanoyl-CoA
2.3 - 385.9
NADH
6 - 90
trans-2-dodecenoyl-CoA
15.3 - 112
trans-2-hexenoyl-CoA
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2000 - 30000
2-dodecenoyl-CoA
15 - 1300
crotonyl-CoA
1300 - 9000
Hexanoyl-CoA
18 - 16000
NADH
390
NADPH
-
wild-type, pH not specified in the publication, temperature not specified in the publication
2000 - 30000
trans-2-dodecenoyl-CoA
1300 - 9000
trans-2-hexenoyl-CoA
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4
butyryl-CoA
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pH and temperature not specified in the publication, 0.5 mM inhibitor, wild-type; wild-type, pH not specified in the publication, temperature not specified in the publication
0.000198
crotonoyl-CoA
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in potassium phosphate buffer pH 6.2, at 30°C
0.05 - 0.08
Hexanoyl-CoA
0.00003 - 0.01
lauroyl-CoA
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.005
-
substrate trans-2-octenoyl-CoA with NADH
0.02
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substrate trans-2-hexadecenoyl-CoA with NADH
0.027
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substrate trans-2-hexadecenoyl-CoA with NADPH
0.078
mitochondria, anaerobic conditions, substrates crotonyl-CoA and NADH
0.086
mitochondria, aerobic conditions, substrates crotonyl-CoA and NADH
0.37
purified recombinant enzyme, substrates trans-hexenoyl-CoA and NADPH
0.66
purified recombinant enzyme, substrates crotonyl-CoA and NADPH
0.79
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in 100 mM PIPES (pH 7.0), at 22°C
1.45
purified recombinant enzyme, substrates trans-hexenoyl-CoA and NADH
1.65
purified recombinant enzyme, substrates crotonyl-CoA and NADH
3.88
pH 6.2, 30°C; purified native enzyme, substrate crotonyl-CoA and NADH
43
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using crotonoyl-CoA and NADH as substrates, at 30°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2 - 7
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less than 50% of maximal activity above pH 7.0 and below pH 5.2
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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PDB
SCOP
CATH
ORGANISM
UNIPROT
Clostridium acetobutylicum (strain ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787)
Clostridium acetobutylicum (strain ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787)
Clostridium acetobutylicum (strain ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787)
Treponema denticola (strain ATCC 35405 / CIP 103919 / DSM 14222)
Treponema denticola (strain ATCC 35405 / CIP 103919 / DSM 14222)
Treponema denticola (strain ATCC 35405 / CIP 103919 / DSM 14222)
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15000
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1 * 15000 + 1 * 25000, SDS-PAGE
25000
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1 * 15000 + 1 * 25000, SDS-PAGE
26000
-
? * 26000, SDS-PAGE
28000
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SDS-PAGE
39000
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gel filtration
44800
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; calculated from cDNA
45700
calculated from cDNA
72000
-
gel filtration
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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1 * 15000 + 1 * 25000, SDS-PAGE
monomer
oligomer
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? * 26000, SDS-PAGE
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
in complex with NAD+, vapor diffusion method; structures of CaTER in apo form at 2.1 A resolution, in complex with NADH at 2.0 A resolution and in complex with NAD+ at 2.7 A resolution
purified and crystallized as an apoenzyme and in a complex form with NADH and triclosan. The crystals of native and complexed FabI diffract to resolutions of 2.6 and 1.8 A, respectively. The crystals both belong to space group P2-1, with unit-cell parameters a = 117.32, b = 155.844, c = 129.448 A , beta = 111.061° for the native enzyme and a = 64.784, b = 107.573, c = 73.517 A, beta = 116.162° for the complex
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crystal structure of tdTer is determined to 2.00 A; to 2.0 A resolution
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in complex with NAD+, vapor diffusion method,; TdTER is crystallized in complex with NAD+ at 3.0 A resolution
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, several months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; native enzyme 1687fold to homogeneity in several steps, recombinant His-tagged enzyme from Escherichia coli by affinity chromatography
Ni-NTA agarose column chromatography
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purified by gel-filtration chromatography on a HiLoad 16/60 superdex 200 column
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using Ni-NTA chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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expressed in Escherichia coli as a His-tagged fusion protein
expressed in Escherichia coli strain BL21(DE3)
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expression in Escherichia coli
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gene inhA, overexpression of stable mutant enzymes in Escherichia coli strain BL21(DE3)
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phage library screening, DNA and amino acid sequence determination and analysis, phylogenetic analysis, functional expression as His-tagged enzyme in Escherichia coli
wild-type and deletion mutant BatG expression in Pseudomonas fluorescens strain BCCM_ID9359, Escherichia coli, and Staphylococcus aureus, confers resistance against kalimantacin/batumin
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F11K
kcat slightly decreased compared to wild-type, Km (NADH) decreased compared to wild-type; the mutant shows 198% activity compared to the wild type enzyme
K244A
inactive; mutant shows no activity
K245A
kcat slightly decreased compared to wild-type, Km (NADH) increased compared to wild-type; the mutant shows 67 activity compared to the wild type enzyme
Y225A
kcat highly decreased compared to wild-type, Km (NADH) increased compared to wild-type; the mutant shows 6.7 activity compared to the wild type enzyme
Y235F
inactive; mutant shows no activity
K165A
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site-directed mutagenesis, mutation of putative catalytic residue
K165Q
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site-directed mutagenesis, mutation of putative catalytic residue
Y158F
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site-directed mutagenesis, mutation of putative catalytic residue, mutant can only be stably expressed in Escherichia coli strain BL21(DE3) if altered to a soluble enzyme form dependent on IPTG induction
F295A
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mutant shows kinetic behaviour relatively similar to wild-type toward substrates crotonyl-CoA and hexanoyl-CoA
I287A
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mutant shows significant decreased kcat values compared to wild-type, mutant exhibit larger increases in catalytic efficiency on the longer hexanoyl-CoA substrate (versus the crotonyl-CoA substrate) of 100 and 17fold compared to that of the wild type (7fold) suggesting suggest these mutations may increase the accessibility of the longer acyl chain to the active site pocket. Mutant shows a much lower Ki (lauroyl-CoA) than wild-type
L276A/V277A
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mutant shows significant decreased kcat values compared to wild-type
L276A/V277A/F295A
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mutant shows significant decreased kcat values compared to wild-type, mutant exhibit larger increases in catalytic efficiency on the longer hexanoyl-CoA substrate (versus the crotonyl-CoA substrate) of 100 and 17fold compared to that of the wild type (7fold) suggesting suggest these mutations may increase the accessibility of the longer acyl chain to the active site pocket
L291A
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mutant shows kinetic behaviour relatively similar to wild-type toward substrates crotonyl-CoA and hexanoyl-CoA
Y240F
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Y240F mutation leads to a 5000fold decrease in catalytic efficiency compared to wild-type with no significant change in Km
Y370A
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mutant shows kinetic behaviour relatively similar to wild-type toward substrates crotonyl-CoA and hexanoyl-CoA
F295A
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mutant shows kinetic behaviour relatively similar to wild-type toward substrates crotonyl-CoA and hexanoyl-CoA
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I287A
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mutant shows significant decreased kcat values compared to wild-type, mutant exhibit larger increases in catalytic efficiency on the longer hexanoyl-CoA substrate (versus the crotonyl-CoA substrate) of 100 and 17fold compared to that of the wild type (7fold) suggesting suggest these mutations may increase the accessibility of the longer acyl chain to the active site pocket. Mutant shows a much lower Ki (lauroyl-CoA) than wild-type
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L291A
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mutant shows kinetic behaviour relatively similar to wild-type toward substrates crotonyl-CoA and hexanoyl-CoA
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Y240F
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Y240F mutation leads to a 5000fold decrease in catalytic efficiency compared to wild-type with no significant change in Km
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Y370A
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mutant shows kinetic behaviour relatively similar to wild-type toward substrates crotonyl-CoA and hexanoyl-CoA
-
additional information
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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enzyme is a target for drugs in treatment of tuberculosis
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Show AA Sequence (405 entries)
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LINKS TO OTHER DATABASES (specific for EC-Number 1.3.1.44)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
InterPro (database of protein families, domains and functional sites)