EC Number | Cloned (Comment) | Organism |
---|---|---|
2.3.1.133 | overexpression of N-terminally His6-tagged HCT in Escherichia coli strain BL21 Star(DE3)pLysS using a commercially available synthetic gene encoding HCT with codons optimized for Escherichia coli expression | Coffea canephora |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
2.3.1.133 | purified recombinant HCT, sitting drop technique, mixing of 100 nl of 20 mg/ml protein in 20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM 2-mercaptoethanol, with 100 nl of precipitant solution containing 1.6 M magnesium sulfate, 0.1 M MES pH 6.5, 20°C, 2 months, X-ray diffraction structure determination and analysis at 3.0 A resolution, molecular replacement | Coffea canephora |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.3.1.133 | 45000 | 50000 | recombinant HCT, gel filtration | Coffea canephora |
2.3.1.133 | 48000 | - |
1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry | Coffea canephora |
2.3.1.133 | 48464 | - |
1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry | Coffea canephora |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.3.1.133 | caffeoyl-CoA + quinate | Coffea canephora | - |
CoA + 5-caffeoylquinate | - |
r | |
2.3.1.133 | caffeoyl-CoA + shikimate | Coffea canephora | - |
CoA + 5-caffeoylshikimate | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.3.1.133 | Coffea canephora | A4ZKE4 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.3.1.133 | recombinant N-terminally His6-tagged HCT from Escherichia coli strain BL21 Star(DE3)pLysS by nickel affinity chromatography, dialysis, and cleavage of the tag by TEV protease, followed by gel filtration | Coffea canephora |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.3.1.133 | caffeoyl-CoA + quinate | - |
Coffea canephora | CoA + 5-caffeoylquinate | - |
r | |
2.3.1.133 | caffeoyl-CoA + shikimate | - |
Coffea canephora | CoA + 5-caffeoylshikimate | - |
r | |
2.3.1.133 | additional information | HCT has a preference for shikimic acid over quinic acid, substrate specifities of the HCT key enzyme, overview | Coffea canephora | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.3.1.133 | monomer | 1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry | Coffea canephora |
2.3.1.133 | More | the enzyme is cleaved into two fragments of 24528 Da and 23906 Da overtime. The protease-cleavage site is localized in the central part of the enzyme, corresponding to a large crossover loop connecting the N- and C-terminal domains in BAHD structures | Coffea canephora |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.3.1.133 | HCT | - |
Coffea canephora |
2.3.1.133 | hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase | - |
Coffea canephora |
EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|---|
2.3.1.133 | additional information | - |
improved stability in sodium phosphate buffer and at higher pH values | Coffea canephora |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.3.1.133 | evolution | HCT belongs to the large plant acyl-CoA-dependent BAHD acyltransferase superfamily | Coffea canephora |
2.3.1.133 | metabolism | the last step in CGA biosynthesis is generally catalysed by a specific hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase, HQT (c.f. EC 2.3.1.99), but it can also be catalysed by the more widely distributed hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase, HCT, substrate specifities of the HCT key enzyme, overview | Coffea canephora |