EC Number | Cloned (Comment) | Organism |
---|---|---|
4.1.2.61 | expression in Escherichia coli BL21 (DE3) | Pseudomonas fluorescens |
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 |
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 |
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 |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.1.2.61 | Pseudomonas fluorescens | O69762 | - |
- |
4.1.2.61 | Pseudomonas fluorescens AN103 | O69762 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.1.2.61 | nickel-affinity chromatography, the histidine tags are removed | Pseudomonas fluorescens |
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 | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.1.2.61 | hexamer | 2 trimers | Pseudomonas fluorescens |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.1.2.61 | HCHL | - |
Pseudomonas fluorescens |
4.1.2.61 | hydroxycinnamoyl-CoA hydratase/lyase | - |
Pseudomonas fluorescens |
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 |