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Literature summary extracted from

  • Lim, S.; Springstead, J.R.; Yu, M.; Bartkowski, W.; Schroeder, I.; Monbouquette, H.G.
    Characterization of a key trifunctional enzyme for aromatic amino acid biosynthesis in Archaeoglobus fulgidus (2009), Extremophiles, 13, 191-198.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
4.2.1.51 L-tyrosine enzyme activation by 70% at 1 mM Archaeoglobus fulgidus
4.2.1.51 Tyr 1 mM, 70% increase in activity Archaeoglobus fulgidus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.12 overexpressed in Escherichia coli Archaeoglobus fulgidus
4.2.1.51 expressed in Escherichia coli (DE3)-RIL, pIS85 expression plasmid Archaeoglobus fulgidus
4.2.1.51 overexpressed in Escherichia coli Archaeoglobus fulgidus
5.4.99.5 overexpressed in Escherichia coli Archaeoglobus fulgidus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.1.12 L-tyrosine 0.5 mM, 50% inhibition Archaeoglobus fulgidus
1.3.1.12 additional information phenylalanine (up to 5 mM) has no effect. Not inhibited by prephenate or NAD+ at concentrations up to 10 and 2 mM, respectively Archaeoglobus fulgidus
4.2.1.51 L-phenylalanine inhibits activity at 5 microM, competitive inhibition Archaeoglobus fulgidus
4.2.1.51 Phe 0.005 mM, 66% inhibition, competitive with prephenate Archaeoglobus fulgidus
5.4.99.5 additional information activity is not affected by both amino acids Phe and Tyr at concentrations up to 0.5 mM Archaeoglobus fulgidus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.12 0.047
-
 NAD+ pH 6.5, 87°C Archaeoglobus fulgidus
1.3.1.12 0.45
-
 prephenate pH 6.5, 87°C Archaeoglobus fulgidus
4.2.1.51 1.3
-
 phenylpyruvate measuring the appearance of phenylpyruvate at 320 nm, performed with 1 mM prephenate in 200 mM Tris buffer Archaeoglobus fulgidus
4.2.1.51 1.3
-
 prephenate 77°C, pH not specified in the publication Archaeoglobus fulgidus
5.4.99.5 additional information
-
 additional information due to instability of chorismate at higher temperature, a Km value is not determined Archaeoglobus fulgidus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.1.12 66000
-
 6 * 66000, SDS-PAGE Archaeoglobus fulgidus
1.3.1.12 70946
-
 6 * 70946, calculated from sequence Archaeoglobus fulgidus
1.3.1.12 420000
-
 gel filtration Archaeoglobus fulgidus
4.2.1.51 66000
-
 trifunctional enzyme, SDS-PAGE Archaeoglobus fulgidus
4.2.1.51 70946
-
 6 * 70946, calculated from sequence Archaeoglobus fulgidus
4.2.1.51 70950
-
 calculated by sequence Archaeoglobus fulgidus
4.2.1.51 420000
-
 gel filtration Archaeoglobus fulgidus
5.4.99.5 66000
-
 6 * 66000, SDS-PAGE Archaeoglobus fulgidus
5.4.99.5 70946
-
 6 * 70946, calculated from sequence Archaeoglobus fulgidus
5.4.99.5 420000
-
 gel filtration Archaeoglobus fulgidus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.1.12 prephenate + NAD+ Archaeoglobus fulgidus the enzyme is involved in aromatic amino acid biosynthesis 4-hydroxyphenylpyruvate + CO2 + NADH
-
 ?
4.2.1.51 prephenate Archaeoglobus fulgidus the enzyme is involved in aromatic amino acid biosynthesis phenylpyruvate + H2O + CO2
-
 ?
5.4.99.5 Chorismate Archaeoglobus fulgidus
-
 Prephenate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.12 Archaeoglobus fulgidus O30012
-
-
4.2.1.51 Archaeoglobus fulgidus O30012
-
-
4.2.1.51 Nanoarchaeum equitans Q74NC4
-
-
5.4.99.5 Archaeoglobus fulgidus O30012
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.12
-
 Archaeoglobus fulgidus
4.2.1.51
-
 Archaeoglobus fulgidus
4.2.1.51 gel filtration Archaeoglobus fulgidus
5.4.99.5
-
 Archaeoglobus fulgidus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.51 additional information
-
 trifunctional enzyme containing prephenate dehydratase (PDT), also chorismate mutase (CM, 5.4.99.5), and prephenate dehydrogenase (PDHG, 1.3.1.12) acitivities Nanoarchaeum equitans
4.2.1.51 additional information
-
 trifunctional enzyme containing prephenate dehydratase (PDT), also chorismate mutase (CM, 5.4.99.5), and prephenate dehydrogenase (PDHG, 1.3.1.12) acitivities, prephenate dehydratase activity spectrophotometric quantified by monitoring the appearance of phenylpyruvate at 320 nm in end-point assays Archaeoglobus fulgidus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.12 prephenate + NAD+ the enzyme is involved in aromatic amino acid biosynthesis Archaeoglobus fulgidus 4-hydroxyphenylpyruvate + CO2 + NADH
-
 ?
1.3.1.12 prephenate + NAD+ activity with NADP+ as coenzyme is about 10% of that with NAD+, suggesting that NAD+ is likely the preferred and physiological coenzyme Archaeoglobus fulgidus 4-hydroxyphenylpyruvate + CO2 + NADH
-
 ?
4.2.1.51 prephenate
-
 Archaeoglobus fulgidus phenylpyruvate + H2O + CO2
-
 ?
4.2.1.51 prephenate prephenate dehydratase part of trifunctional enzyme Nanoarchaeum equitans phenylpyruvate + H2O + CO2
-
 ?
4.2.1.51 prephenate spectrophotometric assay by measuring the appearance of phenylpyruvate Archaeoglobus fulgidus phenylpyruvate + H2O + CO2
-
 ?
4.2.1.51 prephenate the enzyme is involved in aromatic amino acid biosynthesis Archaeoglobus fulgidus phenylpyruvate + H2O + CO2
-
 ?
5.4.99.5 Chorismate
-
 Archaeoglobus fulgidus Prephenate
-
 ?
5.4.99.5 Chorismate the enzyme is involved in aromatic amino acid biosynthesis Archaeoglobus fulgidus Prephenate
-
 ?

Subunits

EC Number Subunits Comment Organism
1.3.1.12 homohexamer 6 * 66000, SDS-PAGE Archaeoglobus fulgidus
1.3.1.12 homohexamer 6 * 70946, calculated from sequence Archaeoglobus fulgidus
4.2.1.51 homohexamer 6 * 66000, SDS-PAGE Archaeoglobus fulgidus
4.2.1.51 homohexamer trifunctional enzyme, SDS-PAGE Archaeoglobus fulgidus
4.2.1.51 homohexamer 6 * 70946, calculated from sequence Archaeoglobus fulgidus
5.4.99.5 homohexamer 6 * 66000, SDS-PAGE Archaeoglobus fulgidus
5.4.99.5 homohexamer 6 * 70946, calculated from sequence Archaeoglobus fulgidus

Synonyms

EC Number Synonyms Comment Organism
1.3.1.12 AroQ gene name. Trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase) Archaeoglobus fulgidus
1.3.1.12 CM/PDT/PDHG trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase) Archaeoglobus fulgidus
1.3.1.12 PDHG
-
 Archaeoglobus fulgidus
4.2.1.51 AroQ gene name. Trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase) Archaeoglobus fulgidus
4.2.1.51 CM/PDT/PDHG trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase) Archaeoglobus fulgidus
4.2.1.51 PDT
-
 Archaeoglobus fulgidus
4.2.1.51 PDT protein
-
 Archaeoglobus fulgidus
4.2.1.51 PDT protein
-
 Nanoarchaeum equitans
5.4.99.5 AroQ gene name. Trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase) Archaeoglobus fulgidus
5.4.99.5 CM/PDT/PDHG trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase) Archaeoglobus fulgidus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.1.12 87
-
 assay at Archaeoglobus fulgidus
4.2.1.51 77
-
 assay at Archaeoglobus fulgidus
4.2.1.51 77
-
 spectrophotometric assay at Archaeoglobus fulgidus
5.4.99.5 60
-
 assay at Archaeoglobus fulgidus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4.99.5 7.5
-
 assay at Archaeoglobus fulgidus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.1.51 5.5 9.5 spectrophotometric assay performed at several different pH values Archaeoglobus fulgidus

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.12 NAD+ activity with NADP+ as coenzyme is about 10% of that with NAD+, suggesting that NAD+ is likely the preferred and physiological coenzyme Archaeoglobus fulgidus

General Information

EC Number General Information Comment Organism
1.3.1.12 metabolism the enzyme is involved in aromatic amino acid biosynthesis Archaeoglobus fulgidus
4.2.1.51 metabolism the enzyme is involved in aromatic amino acid biosynthesis Archaeoglobus fulgidus
5.4.99.5 metabolism the enzyme is involved in aromatic amino acid biosynthesis Archaeoglobus fulgidus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.3.1.12 880
-
 prephenate pH 6,5, 87°C Archaeoglobus fulgidus
4.2.1.51 0.78
-
 prephenate 77°C, pH not specified in the publication Archaeoglobus fulgidus