Information on EC 4.2.1.101 - trans-feruloyl-CoA hydratase

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

EC NUMBER
COMMENTARY hide
4.2.1.101
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RECOMMENDED NAME
GeneOntology No.
trans-feruloyl-CoA hydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-hydroxy-3-methoxyphenyl-beta-hydroxypropanoyl-CoA = feruloyl-CoA + H2O
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cleavage of a C-O bond
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
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capsaicin biosynthesis
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ferulate degradation
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Metabolic pathways
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Phenylalanine metabolism
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SYSTEMATIC NAME
IUBMB Comments
4-hydroxy-3-methoxyphenyl-beta-hydroxypropanoyl-CoA hydro-lyase (feruloyl-CoA-forming)
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CAS REGISTRY NUMBER
COMMENTARY hide
197462-62-7
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
transgenic tobacco cv XHFD 8 expressing various levels of HCHL, the bacterial 4-hydroxycinnamoyl-CoA hydratase/lyase gene of Pseudomonas fluorescens, transgenic lines 201 and 27 reveal high expression, line 13 shows intermediate expression, pBin 19 plants serve as controls
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Manually annotated by BRENDA team
strain HR 199
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Manually annotated by BRENDA team
strain HR 199
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Manually annotated by BRENDA team
gene ferB
UniProt
Manually annotated by BRENDA team
gene ferB
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxycinnamoyl-CoA + H2O
3,4-dihydroxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-caffeoyl-CoA
3,4-dihydroxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-coumaroyl-CoA
4-hydroxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-feruloyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
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i.e. 4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
i.e. vanillin
ir
4-hydroxy-3-methoxycinnamoyl-CoA + H2O
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
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?
4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-hydroxycinnamoyl-CoA + H2O
4-hydroxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-trans-feruloyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
feruloyl-CoA
4-hydroxy-3-methoxy-benzaldehyde + acetyl-CoA
show the reaction diagram
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?
feruloyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
feruloyl-CoA + H2O
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
feruloyl-CoA + H2O
4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
show the reaction diagram
p-coumaroyl-CoA
4-hydroxy-3-methoxy-benzaldehyde + acetyl-CoA
show the reaction diagram
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in transgenic plants the HCHL enzyme acts upon intermediates of the phenylpropanoid pathway
i.e. vanillin
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?
trans-feruloyl-CoA + H2O
4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
show the reaction diagram
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
4-caffeoyl-CoA
3,4-dihydroxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-coumaroyl-CoA
4-hydroxybenzaldehyde + acetyl-CoA
show the reaction diagram
4-feruloyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
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i.e. 4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
i.e. vanillin
ir
4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
feruloyl-CoA
4-hydroxy-3-methoxy-benzaldehyde + acetyl-CoA
show the reaction diagram
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?
feruloyl-CoA
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
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?
feruloyl-CoA + H2O
4-hydroxy-3-methoxybenzaldehyde + acetyl-CoA
show the reaction diagram
trans-feruloyl-CoA + H2O
4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
show the reaction diagram
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.2
4-Coumaroyl-CoA
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1.6
caffeoyl-CoA
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0.0118 - 2.4
Feruloyl-CoA
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.3
4-Coumaroyl-CoA
Pseudomonas fluorescens
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at 30°C
0.41 - 3.72
Feruloyl-CoA
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5 - 9.5
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in Tris-HCl or borate buffer, activity declined to ca. 50% of its maximum at pH 6.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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young leaves from pre- and postflowering transgenic and control plants, cell wall components analysed by HPLC, plants expressing high levels of HCHL exhibit a reduced lignin deposition, monolignol biosynthesis and vascular integrity impaired
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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impact on cell wall integrity and vascular integrity, biomechanical tissue properties, carbon allocation to phenolic pools, lignin formation studied, morphological changes of the plants described
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
31007
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6 * 31007, crystallization, calculated from amino acid sequence
63000
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gel filtration
186000
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calculated from crystallization data
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
additional information
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hexameric structure of two stacked trimers assumed, similarities to the crotonase superfamily
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1.8 A resolution, data collection and refinement statistics shown, structure solution and in silico modelling determined, monomer folding and factors affecting trimer assembly indicated, oligo structure and modelling of the active site determined
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by the hanging-drop method of vapour diffusion using polyethylene glycol 20000 Da as the precipitant, 1.8 A resolution
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hanging-drop vapour diffusion method, wild-type hydroxycinnamoyl-CoA hydratase/lyase in complex with acetyl-CoA and vanillin 2vss: protein concentration of 10 mg/ml in 11% (w/v) PEG [poly(ethylene glycol)] (20000 Da) with 8% PEG (550 Da) monomethyl ether, 0.8 M sodium formate and 0.2% (v/v) butane 1,4-diol in 0.05 M Mes buffer (pH 5.6), 10 mM feruloyl.CoA, resolution 2.22 A, space group P 21 21 21. A ternary complex of hydroxycinnamoyl-CoA hydratase/lyase mutant S123A (HCHL) with acetyl-CoA and vanillin gives insights into substrate specifity and mechanism 2vsu: resolution 1.9 A, space group P 21 21 21
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, several months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Mono-Q, Mono-P, Phenyl-Superose
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nickel-affinity chromatography, the histidine tags are removed
recombinant protein from Escherichia coli
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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expressed in Escherichia coli strain BL21 to yield HCHL protein encoded by Pseudomonas fluorescens strain AN103
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expression in Escherichia coli BL21 (DE3)
expression in Escherichia coli strain BL21(DE3)
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functional expression in hairy root cells of Datura stramonium via infection with Agrobacterium rhizogenes, no accumulation of 4-hydroxybenzaldehyde derivatives occurs, but diminished feruloyl-CoA availability, leading to reduced lignin formation, and formation of new glucoconjugates of 4-hydroxybenzaldehyde derivatives, e.g. 4-hydroxybenzyl alcohol-O-beta-D-glucoside
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gene ferB, the ferBA genes form an operon, genetic organization of the fer gene cluster, overview. Transcriptional regulation of ferBA controlled by a MarR-type transcriptional regulator, FerC. The ferC gene is located upstream of ferB. The transcription start site of ferB is mapped at 30 nucleotides upstream from the ferB initiation codon. Constitutive expression of the ferC-ferB intergenic region in SME043 cells, FerC regulates the ferBA transcription through the binding to the ferC–ferB intergenic region
transgenic tobacco cv. XHFD 8 plants expressing various levels of bacterial HCHL, lignin deposition reduced, monolignol biosynthesis and vascular integrity impaired, reduction in stem toughness, cell wall esterified and soluble phenolics reduced, wall-bound accretion of vanillin and vanillic acid, intracellular accumulation of novel metabolites such as hydroxybenzoic and vanillic acid derivatives
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
transcription of ferB is induced 6.5fold in the SYK-6 cells grown on ferulate
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E143A
Glu143 is essential for catalysis
S123A
mutation of Ser123 to alanine results in a mutant displaying a reduced turnover nummber compared with the wild-type and a KM comparable with that of the wild-type, suggesting that there is little contribution of this residue to mechanism or substrate binding
S123A/E143A
no activity
Y239F
Tyr239 is an important residue in determining substrate specificity
E143A
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Glu143 is essential for catalysis
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S123A
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mutation of Ser123 to alanine results in a mutant displaying a reduced turnover nummber compared with the wild-type and a KM comparable with that of the wild-type, suggesting that there is little contribution of this residue to mechanism or substrate binding
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S123A/E143A
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no activity
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Y239F
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Tyr239 is an important residue in determining substrate specificity
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information