O69762 · HCHL_PSEFL
- ProteinHydroxycinnamoyl-CoA hydratase-lyase
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids276 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Catalyzes the hydration of the acyl-CoA thioester of ferulic acid and the subsequent retro-aldol cleavage of the hydrated intermediate to yield vanillin (4-hydroxy-3-methoxy-benzaldehyde) (PubMed:10222033, PubMed:9461612).
The enzyme is also active with caffeoyl-CoA and 4-coumaroyl-CoA producing 3,4-dihydroxybenzaldehyde and 4-hydroxybenzaldehyde, respectively (PubMed:10222033).
The enzyme is also active with caffeoyl-CoA and 4-coumaroyl-CoA producing 3,4-dihydroxybenzaldehyde and 4-hydroxybenzaldehyde, respectively (PubMed:10222033).
Catalytic activity
- (E)-feruloyl-CoA + H2O = acetyl-CoA + vanillinThis reaction proceeds in the forward direction.
- 3-hydroxy-3-(4-hydroxy-3-methoxyphenyl)propanoyl-CoA = acetyl-CoA + vanillinThis reaction proceeds in the forward direction.
- (E)-caffeoyl-CoA + H2O = 3-hydroxy-3-(3,4-dihydroxyphenyl)propanoyl-CoA
- 3-hydroxy-3-(3,4-dihydroxyphenyl)propanoyl-CoA = 3,4-dihydroxybenzaldehyde + acetyl-CoA
- (E)-4-coumaroyl-CoA + H2O = 3-hydroxy-3-(4-hydroxyphenyl)propanoyl-CoA
- 3-hydroxy-3-(4-hydroxyphenyl)propanoyl-CoA = 4-hydroxybenzaldehyde + acetyl-CoA
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
11.8 μM | feruloyl-CoA | |||||
2.4 μM | feruloyl-CoA | |||||
5.3 μM | 4-coumaroyl-CoA | |||||
1.6 μM | caffeoyl-CoA |
Vmax | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|
36.5 nmol/sec/mg | with feruloyl-CoA as substrate | ||||
73.3 nmol/sec/mg | with 4-coumaroyl-CoA as substrate | ||||
15.2 nmol/sec/mg | with caffeoyl-CoA as substrate |
kcat is 3.72 sec-1 with feruloyl-CoA as substrate.
pH Dependence
Optimum pH is 8.5-9.5.
Features
Showing features for binding site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 29 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: K | ||||||
Binding site | 68 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: A | ||||||
Binding site | 70 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: M | ||||||
Binding site | 72 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: L | ||||||
Binding site | 75 | vanillin (UniProtKB | ChEBI) | ||||
Sequence: Y | ||||||
Binding site | 120 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: G | ||||||
Binding site | 142 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 146 | acetyl-CoA (UniProtKB | ChEBI) | ||||
Sequence: W | ||||||
Binding site | 151 | vanillin (UniProtKB | ChEBI) | ||||
Sequence: G | ||||||
Binding site | 239 | vanillin (UniProtKB | ChEBI) | ||||
Sequence: Y |
GO annotations
Aspect | Term | |
---|---|---|
Molecular Function | feruloyl-CoA hydratase/lyase activity |
Keywords
- Molecular function
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameHydroxycinnamoyl-CoA hydratase-lyase
- EC number
- Short namesHCHL
- Alternative names
Organism names
- Organism
- Strain
- Taxonomic lineageBacteria > Pseudomonadota > Gammaproteobacteria > Pseudomonadales > Pseudomonadaceae > Pseudomonas
Accessions
- Primary accessionO69762
Phenotypes & Variants
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Mutagenesis | 123 | Reduced kcat compared to wild-type but not markerdly. | ||||
Sequence: S → A | ||||||
Mutagenesis | 143 | Abolishes catalytic activity. | ||||
Sequence: E → A | ||||||
Mutagenesis | 239 | Increased KM for feruloyl-CoA but retains a significant amount of catalytic activity with a kcat 10 times less than that of the wild-type. | ||||
Sequence: Y → F |
PTM/Processing
Features
Showing features for initiator methionine, chain.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Initiator methionine | 1 | Removed | ||||
Sequence: M | ||||||
Chain | PRO_0000418727 | 2-276 | Hydroxycinnamoyl-CoA hydratase-lyase | |||
Sequence: STYEGRWKTVKVEIEDGIAFVILNRPEKRNAMSPTLNREMIDVLETLEQDPAAGVLVLTGAGEAWTAGMDLKEYFREVDAGPEILQEKIRREASQWQWKLLRMYAKPTIAMVNGWCFGGGFSPLVACDLAICADEATFGLSEINWGIPPGNLVSKAMADTVGHRQSLYYIMTGKTFGGQKAAEMGLVNESVPLAQLREVTIELARNLLEKNPVVLRAAKHGFKRCRELTWEQNEDYLYAKLDQSRLLDTEGGREQGMKQFLDDKSIKPGLQAYKR |
Interaction
Subunit
Homohexamer; dimer of trimers.
Structure
Sequence
- Sequence statusComplete
- Length276
- Mass (Da)31,008
- Last updated1998-08-01 v1
- ChecksumFDB5AD1539CA7F9A
Features
Showing features for sequence conflict.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Sequence conflict | 16 | in Ref. 1; AA sequence | ||||
Sequence: E → Q |
Keywords
- Technical term