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Literature summary extracted from

  • Bennett, J.P.; Bertin, L.; Moulton, B.; Fairlamb, I.J.; Brzozowski, A.M.; Walton, N.J.; Grogan, G.
    A ternary complex of hydroxycinnamoyl-CoA hydratase-lyase (HCHL) with acetyl-CoA and vanillin gives insights into substrate specificity and mechanism (2008), Biochem. J., 414, 281-289.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.2.61 expression in Escherichia coli BL21 (DE3) Pseudomonas fluorescens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.1.2.61 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 specificity and mechanism 2vsu: resolution 1.9 A, space group P 21 21 21 Pseudomonas fluorescens

Protein Variants

EC Number Protein Variants Comment Organism
4.1.2.61 E143A Glu143 is essential for catalysis Pseudomonas fluorescens
4.1.2.61 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 Pseudomonas fluorescens
4.1.2.61 S123A/E143A no activity Pseudomonas fluorescens
4.1.2.61 Y239F Tyr239 is an important residue in determining substrate specificity Pseudomonas fluorescens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.2.61 additional information
-
 additional information no activity for mutant enzymes E143A and S123A/E143A Pseudomonas fluorescens
4.1.2.61 0.0118
-
 feruloyl-CoA wild-type Pseudomonas fluorescens
4.1.2.61 0.0135
-
 feruloyl-CoA mutant enzyme S123A Pseudomonas fluorescens
4.1.2.61 0.095
-
 feruloyl-CoA mutant enzyme Y239F Pseudomonas fluorescens

Organism

EC Number Organism UniProt Comment Textmining
4.1.2.61 Pseudomonas fluorescens O69762
-
-
4.1.2.61 Pseudomonas fluorescens AN103 O69762
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.2.61 nickel-affinity chromatography, the histidine tags are removed Pseudomonas fluorescens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.2.61 feruloyl-CoA + H2O overall reaction Pseudomonas fluorescens vanillin + acetyl-CoA
-
 ?
4.1.2.61 feruloyl-CoA + H2O overall reaction Pseudomonas fluorescens AN103 vanillin + acetyl-CoA
-
 ?
4.1.2.61 feruloyl-CoA + H2O first step of reaction Pseudomonas fluorescens 4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
-
 ?
4.1.2.61 feruloyl-CoA + H2O first step of reaction Pseudomonas fluorescens AN103 4-hydroxy-3-methoxyphenyl-beta-hydroxypropionyl-CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
4.1.2.61 hexamer 2 trimers Pseudomonas fluorescens

Synonyms

EC Number Synonyms Comment Organism
4.1.2.61 HCHL
-
 Pseudomonas fluorescens
4.1.2.61 hydroxycinnamoyl-CoA hydratase/lyase
-
 Pseudomonas fluorescens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.2.61 additional information
-
 additional information no activity for mutant enzymes E143A and S123A/E143A Pseudomonas fluorescens
4.1.2.61 0.41
-
 feruloyl-CoA mutant enzyme Y239F Pseudomonas fluorescens
4.1.2.61 0.96
-
 feruloyl-CoA mutant enzyme S123A Pseudomonas fluorescens
4.1.2.61 3.72
-
 feruloyl-CoA wild-type Pseudomonas fluorescens