Information on EC 2.3.1.133 - shikimate O-hydroxycinnamoyltransferase

Substrates: -
Products: -

r

caffeoyl-CoA + shikimate

CoA + 5-caffeoylshikimate

Substrates: -
Products: -

r

additional information

?

-

5 entries

Ca2+

Mg2+

2,3-Dihydroxybenzoate

2,5-Dihydroxybenzoate

-

3,4-dihydroxybenzoate

-

3-aminobenzoate

0.8 mM, about 45% inhibition

3-aminobenzoic acid

-

3-Hydroxyanthranilate

Panicum virgatum

R9RYW2

0.8 mM, about 45% inhibition

3-Hydroxybenzoic acid

-

4,4(-diisothiocyano-2,2)-stilbene disulfonic acid

-

486282

4-methyl-1,2,3-thiadiazole-5-carboxylic

-

-

acibenzolar acid

-

Ca2+

diethylpyrocarbonate

-

80% reversion after treatment with hydroxylamine

486282

DTT

Mg2+

p-chloromercuribenzenesulfonic acid

-

protocatechuate

0.8 mM, about 40% inhibition

RNAi

Pinus radiata

-

HCT silencing is achieved by transformation of Pinus radiata callus cells with pHF5

689750

-

Sodium diphosphate

-

tricine

-

weak, not Tris/HCl

additional information

-

no effect: 2,3-butanedione, phenylmethylsulfonylfluoride and N-methylmaleimide

486282

-

EDTA

Tris/HCl

-

activation

0.092 - 0.699

2,3-Dihydroxybenzoate

0.44

2,5-Dihydroxybenzoate

-

pH 7.0, 30°C

1.48

3,4-dihydroxybenzoate

Panicum virgatum

R9RYW2

pH 7.5, 30°C

2.404

3-aminobenzoate

pH 7.0, 30°C, with 4-coumaroyl-CoA

0.366

3-dehydroshikimate

-

pH 7.0, 30°C

0.449 - 0.83

3-Hydroxyanthranilate

0.733 - 1.677

3-hydroxybenzoate

1.1

3-phenylacryloyl-CoA

-

0.0015 - 14.3

4-coumaroyl-CoA

20 entries

0.119

4-coumaroylshikimate

-

0.227 - 0.265

5-O-caffeoylquinate

0.0051 - 30.5

caffeoyl-CoA

14 entries

0.0265

cinnamoyl-CoA

pH 7.0, 30°C, with shikimate

0.019 - 0.162

CoA

0.0051 - 55.5

feruloyl-CoA

8 entries

0.0859

glycerol

Oryza sativa

-

pH 7.5, 37°C, with feruloyl-CoA

720638

0.00582 - 5.82

quinate

8 entries

0.0009 - 83

shikimate

25 entries

0.02

sinapoyl-CoA

pH 7.0, 30°C, with shikimate

additional information

-

0.092

2,3-Dihydroxybenzoate

-

pH 7.0, 30°C

0.44

2,5-Dihydroxybenzoate

-

pH 7.0, 30°C

0.031

3,4-dihydroxybenzoate

-

pH 7.0, 30°C

0.194

3-aminobenzoic acid

-

pH 7.0, 30°C

0.075

3-Hydroxybenzoic acid

-

pH 7.0, 30°C

0.458

4-methyl-1,2,3-thiadiazole-5-carboxylic

-

pH 7.0, 30°C

-

0.216 - 0.254

acibenzolar acid

-

0.0056

-

0.0703

substrates feruloyl-CoA and shikimate, pH 6.5, 30°C

0.0715

Marchantia paleacea

A0A346FEE5

substrates caffeoyl-CoA and shikimate, pH 6.5, 30°C

0.073

substrates feruloyl-CoA and shikimate, pH 6.5, 30°C

0.0736

substrates caffeoyl-CoA and shikimate, pH 6.5, 30°C

0.0807

Marchantia paleacea

A0A346FEE5

substrates 4-coumaroyl-CoA and shikimate, pH 6.5, 30°C

0.0809

substrates 4-coumaroyl-CoA and shikimate, pH 6.5, 30°C

0.0888

substrates 4-coumaroyl-CoA and quinate, pH 6.5, 30°C

0.0903

Marchantia paleacea

A0A346FEE5

substrates 4-coumaroyl-CoA and quinate, pH 6.5, 30°C

0.732

-

1.134

-

additional information

6 entries

6.5

6 entries

6.8

Raphanus sativus

-

50 mM potassium phosphate buffer

486281

7

4 entries

7.1

-

200 mM potassium phosphate buffer

7.5

5 entries

8

-

assay at

additional information

-

pI: 4.63

5 - 9

5.5 - 7.5

-

about half-maximal activity at pH 5.5 and 7.5

6.1 - 6.7

optimal pH range with malate as the acceptor, highest reaction rates are observed for both hydroxycinnamoyl-CoA derivatives between pH 6.1 and 6.7 using sodium phosphate buffer

706306

6.2 - 8.5

-

about half-maximal activity at pH 6.2 in potassium phosphate buffer and 8.5 in Tris-HCl puffer

6.3 - 7.7

-

about 80% of maximal activity at pH 6.3 and 7.7

7.5 - 7.9

optimal pH range with shikimic acid as the acceptor, highest reaction rates are observed for both hydroxycinnamoyl-CoA derivatives between pH 7.5 and 7.9 using sodium phosphate buffer

30

37

40

additional information

Ocimum basilicum

-

above 40°C rapid and irreversible loss of activity

486284

5.78

Brassica oleracea var. oleracea

sequence calculation

-

769950

A0A0D3B2X8

UniProt

Cestrum elegans

D.F.L. von Schlechtendal

-

-

-

-

UniProt

Cynara cardunculus var. scolymus

gene cbhct2

UniProt

-

-

-

UniProt

-

UniProt

-

-

Lithocarpus polystachyus

-

-

-

UniProt

-

SwissProt

SW and EMX-1

-

gene OsHCT4

-

Oryza sativa Japonica Group

-

-

-

UniProt

young green dates

-

-

-

-

UniProt

-

-

-

-

-

-

rye, var. Kustro

-

-

UniProt

-

-

-

-

cv. Compadre

-

cell suspension culture

-

A0A4D6ZMT8, A0A482LP01

-

-

-

-

-

additional information

cytoplasm

Taraxacum antungense

A0A4D6ZMT8, A0A482LP01

-

A0A4D6ZMT8, A0A482LP01

-

5654

757945

additional information

Highest Expressing Human Cell Lines

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Cell Line Links

Gene Links

evolution

5 entries

malfunction

metabolism

4 entries

physiological function

7 entries

additional information

-

PeHCT contains a conserved HXXXDG motif that is found in acyltransferase

Selaginella moellendorffii

40000

45000 - 50000

recombinant HCT, gel filtration

47100

x * 47100, about, sequence calculation

48000

48300

calculated from cDNA

706306

48464

1 * 48000, recombinant enzyme, SDS-PAGE, 1 * 48464, mass spectrometry

49000

static light scattering

50000

58000

-

gel filtration

70000

-

x * 70000, about, recombinant GST-tagged HCT, SDS-PAGE

?

4 entries

monomer

additional information

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

in complex with CoA and 4-coumaroyl-shikimate. The 4-coumaroyl-shikimate contacts the enzyme via the phenolic group and carbonyl group of the 4-coumaroyl portion. The shikimate portion contacts the enzyme through both the carboxyl and hydroxyl groups. In the protocatechuate ternary complex, protocatechuate binds in a very similar manner to shikimate, with the carboxyl group making a tight salt bridge with Arg369, and the C3 hydroxyl group interacting with the nitrogen NE2 of His163

Panicum virgatum

R9RYW2

structures in its apo-form and ternary complex with shikimate and 4-coumaroyl-CoA, which is converted to its product during crystal soaking. Residues threonine36, serine38, tyrosine40, histidine162, arginine371, and threonine384 are involved in catalysis and specificity. Histidine162 and threonine36 play a role in the catalytic mechanism. Substrate binding should occur sequentially, with 4-coumaroyl-CoA binding prior to the acyl acceptor molecule. Comparison of the structure of sorghum HCT with the HCT involved in chlorogenic acid synthesis in Coffea canephora

H162A

complete loss of activity

R371A

complete loss of activity

S38A

40% of wild-type activity

T36A

6% of wild-type activity

T384A

9% of wild-type activity

Y40A

62% of wild-type activity

additional information

80

-

denaturation after 5 min

glycerol stabilizes during storage

-

-20°C, 0.1 M phosphate buffer, pH 6.5, 65% of initial activity retained for at least 85 days

-

-20°C, at least 6 months

-

-20°C, for several months with 10-20% loss of apparent activities after thawing

-

-20°C, for several months, no apparent loss of activity

-

486281

-20°C, with 10% glycerol at least 30 days

-

-80°C, for several months with 10-20% loss of apparent activities after thawing

-

incubation for days at room temperature, multiple freeze/thaw cycles, holding at -20°C for several months, with or without the addition of 10% glycerol, no loss of activity

Ocimum basilicum

-

486284

-

crude extracts from stem material from various down-regulated and control lines are assayed for extractable HCT enzyme activity

-

partial

recombinant enzyme

recombinant GST-tagged HCT from Escherichia coli strain BL21 (DE3) by glutathione affinity chromatography

-

recombinant GST-tagged PeHCT-1, PeHCT-2, PeHCT-3, PeHCT-4, PeHCT-5, PeHCT-6, and PeHCT-7 gene products from Escherichia coli strain BL21 (DE3) by glutathione affinity cromatography

-

recombinant His-tagged HST from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography and dialysis

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

soluble recombinant GST-tagged OsHCT4 from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography

Oryza sativa

-

720638

tracheary elements are cultured

-

-

an antisense construct is generated for down-regulation of HCT

Medicago sativa

-

689416

coding sequences of ZjHCT4 are recombined into the 3302YUBI vector (UBI::ZjHCT4) which are then transformed into Agrobacterium tumefaciens strain EHA105. Expression in Agrostis stolonifera causes excessive elongation and lignin composition changes

Zoysia japonica

-

777772

expressed in Escherichia coli

706306

expression in Escherichia coli

6 entries

expression of GST-tagged HCT in Escherichia coli strain BL21 (DE3)

-

gene cbhct2, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of His-tagged HST in Escherichia coli strain BL21(DE3)pLysS

gene OsHCT4, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression as soluble GST-tagged enzyme using vector pGEX 5X-1 in Escherichia coli strain BL21(DE3)

Oryza sativa

-

720638

genes PeHCT-1, PeHCT-2, PeHCT-3, PeHCT-4, PeHCT-5, PeHCT-6, and PeHCT-7, expression as GST-tagged enzymes in Escherichia coli strain BL21 (DE3)

-

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