Information on EC 4.2.1.119 - enoyl-CoA hydratase 2

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

EC NUMBER
COMMENTARY hide
4.2.1.119
-
RECOMMENDED NAME
GeneOntology No.
enoyl-CoA hydratase 2
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(3R)-3-hydroxyacyl-CoA = (2E)-2-enoyl-CoA + H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydration
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
10-cis-heptadecenoyl-CoA degradation (yeast)
-
-
10-trans-heptadecenoyl-CoA degradation (MFE-dependent, yeast)
-
-
10-trans-heptadecenoyl-CoA degradation (reductase-dependent, yeast)
-
-
6-gingerol analog biosynthesis (engineered)
-
-
9-cis, 11-trans-octadecadienoyl-CoA degradation (isomerase-dependent, yeast)
-
-
fatty acid beta-oxidation (peroxisome, yeast)
-
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fatty acid beta-oxidation VI (peroxisome)
-
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polyhydroxydecanoate biosynthesis
-
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stearate biosynthesis I (animals and fungi)
-
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unsaturated, even numbered fatty acid beta-oxidation
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SYSTEMATIC NAME
IUBMB Comments
(3R)-3-hydroxyacyl-CoA hydro-lyase
This enzyme catalyses a hydration step in peroxisomal beta-oxidation. The human multifunctional enzyme type 2 (MFE-2) is a 79000 Da enzyme composed of three functional units: (3R)-hydroxyacyl-CoA dehydrogenase, 2-enoyl-CoA hydratase 2 and sterol carrier protein 2-like units [1]. The enzymes from Aeromonas caviae [4] and Arabidopsis thaliana [5] are monofunctional enzymes. 2-Enoyl-CoA hydratase 3 from Candida tropicalis is a part from multifunctional enzyme type 2 [3].
CAS REGISTRY NUMBER
COMMENTARY hide
9027-13-8
cf. EC 4.2.1.17
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
L. cv. Chinesische Schlangengurken
-
-
Manually annotated by BRENDA team
presence of 16 orthologues of R-specific enoyl-CoA hydratase, among which three proteins shares high homologies with the enzyme specific to enoyl-CoAs of medium chain length encoded by phaJ4 from Pseudomonas aeruginosa
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-
Manually annotated by BRENDA team
presence of 16 orthologues of R-specific enoyl-CoA hydratase, among which three proteins shares high homologies with the enzyme specific to enoyl-CoAs of medium chain length encoded by phaJ4 from Pseudomonas aeruginosa
-
-
Manually annotated by BRENDA team
gene CG3415
-
-
Manually annotated by BRENDA team
the classification is ambiguous because the stereochemistry of the reaction product is not exactly determined
-
-
Manually annotated by BRENDA team
fox2 mutant strain
UniProt
Manually annotated by BRENDA team
fox2 mutant strain
UniProt
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
(24E)-3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-24-enoyl-CoA + H2O
(24R,25R)-3alpha,7alpha,12alpha,24-tetrahydroxy-5beta-cholestanoyl-CoA
show the reaction diagram
-
reaction of the recombinant enzyme, protein converted rapidly
a physiological intermediate in bile acid synthesis
-
?
(2E)-2-decenoyl-CoA + H2O
(3R)-3-hydroxydecanoyl-CoA
show the reaction diagram
-
activity measurements are based on the formation of the magnesium complex of 3-ketoacyl-CoA from (2E)-2-decenoyl-CoA
-
-
?
(2E)-2-enoyl-CoA + H2O
(3R)-3-hydroxyacyl-CoA
show the reaction diagram
(2E)-butenoyl-CoA + H2O
(3R)-hydroxybutanoyl-CoA
show the reaction diagram
-
-
-
-
?
(2E)-decenoyl-CoA + H2O
(3R)-3-hydroxydecanoyl-CoA
show the reaction diagram
(2E)-enoyl-CoA + H2O
(3R)-hydroxyacyl-CoA
show the reaction diagram
(2E)-hexadecenoyl-CoA + H2O
(3R)-3-hydroxyhexadecanoyl-CoA
show the reaction diagram
-
-
-
?
(2E)-hexenoyl-CoA + H2O
(3R)-3-hydroxyhexanoyl-CoA
show the reaction diagram
(3R)-3-hydroxyacyl-CoA
(2E)-2-enoyl-CoA + H2O
show the reaction diagram
(3R)-3-hydroxydecanoyl-CoA
(2E)-2-decenoyl-CoA + H2O
show the reaction diagram
(3R)-3-hydroxyhexadecanoyl-CoA
(2E)-2-hexadecenoyl-CoA + H2O
show the reaction diagram
(R)-3-hydroxydecanoyl-CoA
trans-2-decenoyl-CoA + H2O
show the reaction diagram
-
-
-
-
r
(R)-3-hydroxyoctanoyl-CoA
octenoyl-CoA + H2O
show the reaction diagram
-
no activity with (S)-3-hydroxyoctanoyl-CoA
-
-
r
2-trans-butenoyl-CoA + H2O
(3R)-hydroxybutanoyl-CoA
show the reaction diagram
-
-
-
-
?
2-trans-decenoyl-CoA + H2O
(3R)-3-hydroxydecanoyl-CoA
show the reaction diagram
-
-
-
-
?
Crotonyl-CoA + H2O
(3R)-3-Hydroxybutanoyl-CoA
show the reaction diagram
crotonyl-CoA + H2O
(R)-3-hydroxybutanoyl-CoA
show the reaction diagram
crotonyl-CoA + H2O
3-hydroxybutanoyl-CoA
show the reaction diagram
-
-
-
?
crotonyl-CoA + H2O
?
show the reaction diagram
dec-2-enoyl-CoA + H2O
(R)-3-hydroxydecanoyl-CoA
show the reaction diagram
-
9-12% of the activity with hexenoyl-CoA, depending on preparation
-
-
?
dec-2-enoyl-CoA + H2O
3-hydroxydecanoyl-CoA
show the reaction diagram
-
-
-
?
dodec-2-enoyl-CoA + H2O
(R)-3-hydroxydodecenoyl-CoA
show the reaction diagram
-
4-5% of the activity with hexenoyl-CoA, depending on preparation
-
-
?
dodec-2-enoyl-CoA + H2O
3-hydroxydodecanoyl-CoA
show the reaction diagram
-
-
-
?
hex-2-enoyl-CoA + H2O
(R)-3-hydroxyhexanoyl-CoA
show the reaction diagram
hexenoyl-CoA + H2O
3-hydroxyhexanoyl-CoA
show the reaction diagram
-
-
-
?
oct-2-enoyl-CoA + H2O
(R)-3-hydroxyoctanoyl-CoA
show the reaction diagram
oct-2-enoyl-CoA + H2O
3-hydroxyoctanoyl-CoA
show the reaction diagram
-
-
-
?
pent-2-enoyl-CoA + H2O
(R)-3-hydroxypentanoyl-CoA
show the reaction diagram
-
-
-
-
?
tetradec-2-enoyl-CoA + H2O
?
show the reaction diagram
-
-
-
-
?
trans-2-decenoyl-CoA
(3R)-hydroxydecanoyl-CoA + H2O
show the reaction diagram
-
-
-
-
?
trans-2-decenoyl-CoA + H2O
(3R)-3-hydroxydecanoyl-CoA
show the reaction diagram
-
-
-
-
r
trans-2-decenoyl-CoA + H2O
(3R)-hydroxydecanoyl-CoA
show the reaction diagram
-
ratio of hydration rates trans-2-decenoyl-CoA/crotonyl-CoA is 14.4
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r
trans-2-hexadecenoyl-CoA
(3R)-hydroxyhexadecanoyl-CoA + H2O
show the reaction diagram
-
-
-
-
?
trans-2-octenoyl-CoA + H2O
3-hydroxyoctanoyl-CoA
show the reaction diagram
-
-
-
?
trans-dec-2-enoyl-CoA
?
show the reaction diagram
-
activity is 7fold lower than activity with crotonyl-CoA
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-
?
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
(2E)-2-enoyl-CoA + H2O
(3R)-3-hydroxyacyl-CoA
show the reaction diagram
(2E)-enoyl-CoA + H2O
(3R)-hydroxyacyl-CoA
show the reaction diagram
-
-
-
-
?
(3R)-3-hydroxyacyl-CoA
(2E)-2-enoyl-CoA + H2O
show the reaction diagram
(3R)-3-hydroxydecanoyl-CoA
(2E)-2-decenoyl-CoA + H2O
show the reaction diagram
-
-
-
-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(R)-methylenecyclopropylformyl-CoA
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methylenecyclopropylformyl-CoA is a better inhibitor for enoyl-CoA hydratase 2 than for enoyl-CoA hydratase 1
(S)-methylenecyclopropylformyl-CoA
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methylenecyclopropylformyl-CoA is a better inhibitor for enoyl-CoA hydratase 2 than for enoyl-CoA hydratase 1
3-octynoyl-CoA
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irreversible inhibitor of enoyl-CoA hydratase 2
NEM
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60% inhibition by 5 mM, 96% inhibition by 10 mM NEM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0081 - 0.0131
(2E)-2-decenoyl-CoA
0.0853
(2E)-butenoyl-CoA
-
pH 7.5, 22°C, recombinant full-length enzyme
0.00114 - 4.6
(2E)-decenoyl-CoA
0.0667 - 8.7
(2E)-hexenoyl-CoA
0.021 - 0.076
(2E)-oct-2-enoyl-CoA
0.11
2-trans-butenoyl-CoA
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pH 8.0, 25°C
-
0.0095
2-trans-decenoyl-CoA
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pH 8.0, 25°C
0.024 - 60
crotonyl-CoA
0.007
dec-2-enoyl-CoA
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-
0.013 - 0.042
decenoyl-CoA
0.005 - 0.043
dodec-2-enoyl-CoA
0.015 - 0.034
hex-2-enoyl-CoA
0.018 - 0.102
hexenoyl-CoA
0.009 - 0.05
oct-2-enoyl-CoA
0.036
pent-2-enoyl-CoA
-
pH 8.0, 30°C
0.005
tetradec-2-enoyl-CoA
-
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.3 - 388
(2E)-2-decenoyl-CoA
0.38
(2E)-butenoyl-CoA
Drosophila melanogaster
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pH 7.5, 22°C, recombinant full-length enzyme
26 - 1100
(2E)-decenoyl-CoA
22.8 - 210
(2E)-hexenoyl-CoA
2.3
crotonyl-CoA
Rattus norvegicus
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-
additional information
additional information
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000446
(2E)-butenoyl-CoA
Drosophila melanogaster
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pH 7.5, 22°C, recombinant full-length enzyme
41716
0.97
(2E)-decenoyl-CoA
Drosophila melanogaster
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pH 7.5, 22°C, recombinant full-length enzyme
14250
0.0032
(2E)-hexenoyl-CoA
Drosophila melanogaster
-
pH 7.5, 22°C, recombinant full-length enzyme
11852
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.041
(R)-methylenecyclopropylformyl-CoA
-
25°C
0.053
(S)-methylenecyclopropylformyl-CoA
-
25°C
0.065
3-octynoyl-CoA
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25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.05
-
L654G/V130G mutant with crotonyl-CoA as substrate, cell extract
0.08
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L65A/V130G mutant with crotonyl-CoA as substrate, cell extract
0.12
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strain HsMFE-2(D490A)
0.16
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strain HsMFE-2(G16S)
0.2
-
strain HsMFE-2(H532A)
0.21
-
strain HsMFE-2(D370A), strain HsMFE-2(H406A)
0.24
-
strain HsMFE-2(Y410A)
0.26
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strain HsMFE-2(D517A), strain HsMFE-2(E408A)
0.4
-
strain UTL-7A
0.54
-
strain HsMFE-2
0.55
-
S62A mutant with octenoyl-CoA as substrate, cell extract
0.86
-
wild-type with octenoyl-CoA as substrate, cell extract
0.994
-
pH and temperature not specified in the publication
1.2
0.03 mM (2E)-hexadecenoyl-CoA as a substrate
1.98
-
L65G mutant with octenoyl-CoA as substrate, cell extract
2.25
-
L65I mutant with octenoyl-CoA as substrate, cell extract
3.5
0.03 m (2E)-hexenoyl-CoA as a substrate
6.59
-
V130A mutant with octenoyl-CoA as substrate, cell extract
7.92
-
L65V mutant with octenoyl-CoA as substrate, cell extract
12.4
0.1 mM, 3-hydroxydecanoyl-CoA as a substrate
15.8
-
L65G mutant with crotonyl-CoA as substrate, cell extract
21.2
-
V130G mutant with octenoyl-CoA as substrate, cell extract
30
0.05 mM (2E)-decenoyl-CoA as a substrate
33.4
-
pET-Hydr2 expressed in Escherichia coli, soluble extract of the cells
37.5
-
purified enzyme
48
-
recombinant 46 kDa hydratase 2, last purification step: size exclusion
67
-
S62A mutant with crotonyl-CoA as substrate, cell extract
68.5
-
V130G mutant with crotonyl-CoA as substrate, cell extract
69.8
-
L65A mutant with octenoyl-CoA as substrate, cell extract
883
-
V130G mutant with crotonyl-CoA as substrate, purified enzyme
1256
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L65A mutant with crotonyl-CoA as substrate, cell extract
1288
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V130A mutant with crotonyl-CoA as substrate, cell extract
1538
-
L65V mutant with crotonyl-CoA as substrate, cell extract
1594
-
wild-type with crotonyl-CoA as substrate, cell extract
1880
-
L65I mutant with crotonyl-CoA as substrate, cell extract
additional information
-
activity is below the detection limit of the assay system when using extracts from non-transformed cells or cells transformed with the vector only
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8
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-
9
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
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pH dependence experiment is performed in 200 mM potassium phosphate buffer at pH values varying from 5 to 10
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6
calculated from sequence
9
-
around, estimated by comparing the elution of cationic enzymes on carboxymethyl-cellulose
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
fat-degrading
Manually annotated by BRENDA team
AtECH2 gene expression is strongest in tissues with high beta-oxidation activity, such as germinating seedlings and senescing leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13954
-
2 * 13954, calculated from sequence
16500
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x * 17500, recombinant isozyme His-tagged PhaJ4aRe, SDS-PAGE, x * 17000, recombinant His-tagged PhaJ4bRe, SDS-PAGE, x * 16500, recombinant His-tagged PhaJ4cRe, SDS-PAGE
17000
-
x * 17500, recombinant isozyme His-tagged PhaJ4aRe, SDS-PAGE, x * 17000, recombinant His-tagged PhaJ4bRe, SDS-PAGE, x * 16500, recombinant His-tagged PhaJ4cRe, SDS-PAGE
17500
-
x * 17500, recombinant isozyme His-tagged PhaJ4aRe, SDS-PAGE, x * 17000, recombinant His-tagged PhaJ4bRe, SDS-PAGE, x * 16500, recombinant His-tagged PhaJ4cRe, SDS-PAGE
30000
-
gel filtration
31500
-
2 * 31500, microsomal isoform, SDS-PAGE
33000
-
2 * 33000, SDS-PAGE
33500
-
2 * 33500, peroxisomal isoform, SDS-PAGE
34000
x * 34000, calculated from sequence
45000
-
x * 45000, HsMFE-2(dhdelta), HsMFE-2(dhdelta, E366A), HsMFE-2(dhdelta, D510A), SDS-PAGE
46000
-
recombinant hydratase 2, SDS-PAGE
60000 - 88000
-
-
60000
-
microsomal isoform, gel filtration
62000
-
peroxisomal isoform, gel filtration
63000
-
both recombinant CtMfe2p(dha+bdelta) and its SeMet analogue, SDS-PAGE
64100
-
2 * 64100, MFE-2, SDS-PAGE
65000
-
gel filtration
80000
-
x * 80000, SDS-PAGE
155000
-
recombinant enzyme, sucrose density gradient centrifugation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
-
size-exclusion chromatography on a Superdex 200 HR column gives a native molecular mass of 59 kDa, suggesting that the recombinant protein is monomeric
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapour diffusion against a reservoir solution containing 20% polyethylene glycol 4000, 5% 2-propanol and 20 mM HEPES pH 7.0 at 25°C. Crystals belong to the monoclinic space group C2, with unit-cell parameters a = 111.54 A, b = 59.29 A, c = 47.27 A, beta = 113.04° and contain a dimeric molecule in the asymmetric unit
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hanging-drop vapour-diffusion method. Crystals of native and SeMet CtMfe2p(dha+bdelta)
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structure determination. The eukaryotic hydratase 2 has a complete hot dog fold only in its C-domain, whereas the N-domain lacks a long central alpha-helix, thus creating space for bulkier substrates in the binding pocket. The hydrogen bonding network of the active site of 2-enoyl-CoA hydratase 2 resembles the active site geometry of mitochondrial (S)-specific 2-enoyl-CoA hydratase 1, although in a mirror image fashion
purified recombinant detagged MFE-2, 5 mg/ml protein in 0.1 Msodium phosphate, pH 7.2, and 0.2 M NaF, sitting and hanging drop vapour diffusion methods are used at 21°C, mixing of equal volumes of protein and reservoir solutions, the latter contains 100 mM Tris-HCl, pH 8.0, 1.0 M NaCl, 20% w/v PEG 5000 MME and 5 mM NAD+, X-ray diffraction structure determination and analysis at 2.15 A resolution
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hanging-drop vapor diffusion method, crystal structure to 3 A resolution. MFE-2 has a two-domain subunit structure with a C-domain complete hot-dog fold housing the active site, and an N-domain incomplete hot-dog fold housing the cavity for the aliphatic acyl part of the substrate molecule. The ability of human hydratase 2 to utilize such bulky compounds which are not physiological substrates for the fungal ortholog, e.g. CoA esters of C26 fatty acids, pristanic acid and di/trihydroxycholestanoic acids, is explained by a large hydrophobic cavity formed upon the movements of the extremely mobile loops I–III in the N-domain. In the unliganded form of human hydratase 2, however, the loop I blocks the entrance of fatty enoyl-CoAs with chain-length above C8
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stable for several months
-
The purified enzyme can be stored as an active enzyme for at least half a year at ­4°C or frozen at -20°C.
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
0.2% yield,the microsomal isoform elutes at 0.4-0.7 MKCl, at low salt concentrations, whereas the peroxisomal isoform elutes at high salt concentrations
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recombinant His-tagged MFE-2 from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography, removal of the His tag
-
recombinant His6-tagged PhaJ4 homologues from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant protein purified from the cell extract to apparent homogeneity by three chromatographic steps on anion-exchange, cation-exchange and size-exclusion columns
-
recombinant wild-type and mutant GFP-tagged hydratase domains of multifunctional enzyme type 2 hydratase from Escherichia coli strain JM109 by anion exchange chromatography
-
the FOX2 gene is overexpressed from a multicopy vector (YEp352) in Saccharomyces cerevisiae and the gene product purified to apparent homogenity. A truncated version of MFP lacking 271 carboxyl-terminal amino acids is also overexpressed and purified
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a truncated version (amino acid residues 318-735) of perMFE-2 is expressed in Escherichia coli BL21(DE3) plysS cells as a recombinant protein
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AtECH2 contains a peroxisome targeting signal at the C-terminal end, is addressed to the peroxisome in Saccharomyces cerevisiae, and a fusion protein between AtECH2 and a fluorescent protein is targeted to peroxisomes in onion cells. To assess the peroxisomal addressing of AtECH2, a fusion protein between an EYFP at the N terminus and AtECH2 at the C terminus is constructed and expressed under the control of a double cauliflower mosaic virus (CaMV) 35 S viral promoter to allow transient expression of the fusion protein in onion cells following biolistic bombardment. The fluorescence is examined by confocal microscopy after 12 h
expressed in Escherichia coli BL21(DE3)
-
expression in Escherichia coli BL21 (DE3)
-
expression in Escherichia coli BL21(DE3)
expression in Pichia pastoris
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expression of wild-type and mutant C-terminally His6-tagged MFE2s in Escherichia coli
-
expression of wild-type and mutant hydratase domains of multifunctional enzyme type 2 hydratase as GFP-tagged protein in Escherichia coli strain JM109
-
gene CG3415, Drosophila melanogaster MFE-2 complements a Saccharomyces cerevisiae MFE-2 deletion strain, functional expression of His-tagged MFE-2 in Escherichia coli strain BL21(DE3) pLysS
-
overexpression of the three His6-tagged PhaJ4 homologues individually in Escherichia coli strain BL21(DE3)
-
wild type (HsMFE-2) and its variants are expressed in Saccharomyces cerevisiae, the recombinant HsMFE-2(dhdelta) and its variants are expressed in Escherichia coli BL21(DE3)pLysS
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L654G/V130G
-
decreased specific activity for crotonyl-CoA
L65A
-
enzyme activity similar to wild-type enzyme
L65A/V130G
-
decreased specific activity for crotonyl-CoA
L65G
-
enzyme activity similar to wild-type enzyme
L65I
-
specific activity with crotonyl-CoA similar to wild-type enzyme
L65V
-
specific activity with crotonyl-CoA similar to wild-type enzyme
S62A
-
decreased specific activity for crotonyl-CoA
V130A
-
specific activity with crotonyl-CoA similar to wild-type enzyme
V130G
-
enzyme activity similar to wild-type enzyme, lower structural stability than wild-type enzyme
A348T
-
site-directed mutagenesis, the mutation does not affect the enzyme
A427V
-
site-directed mutagenesis, the mutation does not affect the enzyme, the mutant shows a slight increase in activity
A491T
-
site-directed mutagenesis, the mutation does not affect the enzyme
A606S
-
site-directed mutagenesis, the mutant shows reduced activity
D370A
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
D490A
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
D510A
-
inactive mutant enzyme
D510Y
-
site-directed mutagenesis, inactive mutant, the mutation disrupts active site architecture
D517A
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
E366A
-
kcat/Km 100times lower than that of the wild type
E366G
-
site-directed mutagenesis, inactive mutant, the mutation disrupts dimerization
E408A
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
G16S
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
G533R
-
site-directed mutagenesis, inactive mutant, the mutation disrupts ligand interaction
H406A
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
H515A
-
inactive mutant enzyme
H532A
-
reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
H532R
-
site-directed mutagenesis, inactive mutant, the mutation disrupts active site architecture
I516T
-
site-directed mutagenesis, the mutation disrupts dimerization, the mutant shows reduced activity
I559V
-
site-directed mutagenesis, the mutation does not affect the enzyme
L405P
-
site-directed mutagenesis, inactive mutant, the mutation disrupts ligand interaction
N457D
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site-directed mutagenesis, the mutant shows reduced activity
N457Y
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site-directed mutagenesis, the mutation disrupts domain folding, the mutant shows reduced activity
P529L
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site-directed mutagenesis, inactive mutant, the mutation disrupts active site architecture
R506C
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site-directed mutagenesis, inactive mutant, the mutation disrupts dimerization
R506H
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site-directed mutagenesis, inactive mutant, the mutation disrupts dimerization
W511R
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site-directed mutagenesis, the mutation does not affect the enzyme
Y347A
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inactive mutant enzyme
Y410A
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reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae
Y505A
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inactive mutant enzyme
R251A
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site-directed mutagenesis, the mutant shows decreased labeling efficiency in photoaffinity labeling of substrate binding sites with photophores compared to the wild-type enzyme, overview
W249A
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site-directed mutagenesis, the mutant shows decreased labeling efficiency in photoaffinity labeling of substrate binding sites with photophores compared to the wild-type enzyme, overview
W249G
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site-directed mutagenesis, the mutant shows decreased labeling efficiency in photoaffinity labeling of substrate binding sites with photophores compared to the wild-type enzyme, overview
R251A
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site-directed mutagenesis, the mutant shows decreased labeling efficiency in photoaffinity labeling of substrate binding sites with photophores compared to the wild-type enzyme, overview
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W249A
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site-directed mutagenesis, the mutant shows decreased labeling efficiency in photoaffinity labeling of substrate binding sites with photophores compared to the wild-type enzyme, overview
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W249G
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site-directed mutagenesis, the mutant shows decreased labeling efficiency in photoaffinity labeling of substrate binding sites with photophores compared to the wild-type enzyme, overview
-
DELTA629-990
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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recombinant 46 kDa hydratase 2 survives in a purified form under storage, thus being the first protein of this type amenable to application as a tool in metabolic studies
Show AA Sequence (232 entries)
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