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Literature summary for 2.3.1.133 extracted from

  • Lallemand, L.A.; McCarthy, J.G.; McSweeney, S.; McCarthy, A.A.
    Purification, crystallization and preliminary X-ray diffraction analysis of a hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT) from Coffea canephora involved in chlorogenic acid biosynthesis (2012), Acta Crystallogr. Sect. F, 68, 824-828.
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

Cloned (Comment) Organism
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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000 50000 recombinant HCT, gel filtration Coffea canephora
48000
-
 1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry Coffea canephora
48464
-
 1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry Coffea canephora

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
caffeoyl-CoA + quinate Coffea canephora
-
 CoA + 5-caffeoylquinate
-
 r
caffeoyl-CoA + shikimate Coffea canephora
-
 CoA + 5-caffeoylshikimate
-
 r

Organism

Organism UniProt Comment Textmining
Coffea canephora A4ZKE4
-
-

Purification (Commentary)

Purification (Comment) Organism
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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
caffeoyl-CoA + quinate
-
 Coffea canephora CoA + 5-caffeoylquinate
-
 r
caffeoyl-CoA + shikimate
-
 Coffea canephora CoA + 5-caffeoylshikimate
-
 r
additional information HCT has a preference for shikimic acid over quinic acid, substrate specifities of the HCT key enzyme, overview Coffea canephora ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry Coffea canephora
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

Synonyms

Synonyms Comment Organism
HCT
-
 Coffea canephora
hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase
-
 Coffea canephora

pH Stability

pH Stability pH Stability Maximum Comment Organism
additional information
-
 improved stability in sodium phosphate buffer and at higher pH values Coffea canephora

General Information

General Information Comment Organism
evolution HCT belongs to the large plant acyl-CoA-dependent BAHD acyltransferase superfamily Coffea canephora
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