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Literature summary extracted from

  • Ohara, K.; Mito, K.; Yazaki, K.
    Homogeneous purification and characterization of LePGT1 - a membrane-bound aromatic substrate prenyltransferase involved in secondary metabolism of Lithospermum erythrorhizon (2013), FEBS J., 280, 2572-2580.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.93 recombinant expression of C-terminally His-tagged LePGT1 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection method Lithospermum erythrorhizon

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.93 D212A site-directed mutagenesis, the mutant shows almost no expression of the recombinant protein Lithospermum erythrorhizon
2.5.1.93 D87A site-directed mutagenesis, the mutant can be be successfully expressed, but is catalytically inactive Lithospermum erythrorhizon
2.5.1.93 additional information the substitution of aspartate with alanine, especially D87 and D212, causes a dramatic decrease in enzyme activity Lithospermum erythrorhizon

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.93 2-hydroxybenzoic acid 10% inhibition Lithospermum erythrorhizon
2.5.1.93 caffeic acid 30% inhibition Lithospermum erythrorhizon
2.5.1.93 catechol 89% inhibition Lithospermum erythrorhizon
2.5.1.93 chlorogenic acid 20% inhibition Lithospermum erythrorhizon
2.5.1.93 homogentisic acid 93% inhibition Lithospermum erythrorhizon
2.5.1.93 hydroquinone 98% inhibition Lithospermum erythrorhizon
2.5.1.93 additional information compounds showing strong competition/inhibition are simple phenols having phenolic OH residues, while carboxylic acid does not seem to influence the enzyme activity. Phenylpropanoid gives no apparent inhibition. No or poor inhibition by benzoate, 2-hydroxybenzoate, 3-hydroxybenzoate, ferulic acid, and 4-coumaric acid Lithospermum erythrorhizon

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.93 additional information
-
 additional information Michaelis-Menten kinetics Lithospermum erythrorhizon
2.5.1.93 0.0138
-
 geranyl diphosphate recombinant enzyme, pH 7.0-7.5, 45°C Lithospermum erythrorhizon
2.5.1.93 0.0184
-
 4-hydroxybenzoate recombinant enzyme, pH 7.0-7.5, 45°C Lithospermum erythrorhizon

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.5.1.93 membrane membrane-bound Lithospermum erythrorhizon 16020
-
2.5.1.93 microsome
-
 Lithospermum erythrorhizon
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.93 Co2+ results in 34.8% of the activity with Mg2+ Lithospermum erythrorhizon
2.5.1.93 Fe2+ results in 15.0% of the activity with Mg2+ Lithospermum erythrorhizon
2.5.1.93 Mg2+ preferred divalent cation, required Lithospermum erythrorhizon
2.5.1.93 Mn2+ results in 8.54% of the activity with Mg2+ Lithospermum erythrorhizon
2.5.1.93 additional information no activity with Ca2+, Cu2+, and Zn2+ Lithospermum erythrorhizon

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.93 29000
-
 x * 29000, recombinant His-tagged enzyme, SDS-PAGE Lithospermum erythrorhizon

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.93 geranyl diphosphate + 4-hydroxybenzoate Lithospermum erythrorhizon
-
 3-geranyl-4-hydroxybenzoate + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.93 Lithospermum erythrorhizon Q8W405 gene PGT-1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.93 recombinant C-terminally His-tagged LePGT1 41.3fold from Spodoptera frugiperda Sf9 cells by improved detergent solubilization with 6 mM sodium deoxycholate, and nickel affinity chromatography Lithospermum erythrorhizon

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.93 0.0665
-
 recombinant enzyme in crude Sf9 cell extract, pH 7.0-7.5, 45°C Lithospermum erythrorhizon
2.5.1.93 4.68
-
 purified recombinant enzyme, pH 7.0-7.5, 45°C Lithospermum erythrorhizon

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.93 geranyl diphosphate + 4-hydroxybenzoate
-
 Lithospermum erythrorhizon 3-geranyl-4-hydroxybenzoate + diphosphate
-
 ?
2.5.1.93 geranyl diphosphate + 4-hydroxybenzoate LePGT1 reveals high specificity for PHB to give 3-geranyl-4-hydroxybenzoic acid (100%) Lithospermum erythrorhizon 3-geranyl-4-hydroxybenzoate + diphosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.93 ? x * 29000, recombinant His-tagged enzyme, SDS-PAGE Lithospermum erythrorhizon
2.5.1.93 More the enzyme contains contains multiple transmembrane alpha-helices Lithospermum erythrorhizon

Synonyms

EC Number Synonyms Comment Organism
2.5.1.93 LePGT1
-
 Lithospermum erythrorhizon
2.5.1.93 p-hydroxybenzoate: geranyltransferase
-
 Lithospermum erythrorhizon
2.5.1.93 PHB geranyltransferase
-
 Lithospermum erythrorhizon

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.93 45
-
 recombinant enzyme Lithospermum erythrorhizon

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.93 7 7.5 recombinant enzyme Lithospermum erythrorhizon

General Information

EC Number General Information Comment Organism
2.5.1.93 additional information LePGT1 is a model membrane-bound aromatic substrate prenyltransferase Lithospermum erythrorhizon
2.5.1.93 physiological function the enzyme is involved in the biosynthesis of a red naphthoquinone pigment, shikonin Lithospermum erythrorhizon