Any feedback?
Information on EC 2.5.1.93 - 4-hydroxybenzoate geranyltransferase and Organism(s) Lithospermum erythrorhizon and UniProt Accession Q8W404
for references in articles please use BRENDA:EC2.5.1.93Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
2.5.1.93 4-hydroxybenzoate geranyltransferaseIUBMB Comments
The enzyme is involved in shikonin biosynthesis. It has a strict substrate specificity for geranyl diphosphate and an absolute requirement for Mg2+ .
Specify your search results
Word Map
- 2.5.1.93
- shikonins
- naphthoquinone
-
lithospermum
- euchroma
- arnebia
- erythrorhizon
-
geranylation
- prenyltransferase
-
hairy
-
royle
- ubiquinone
- boraginaceae
- acetylshikonin
- mevalonate
-
phenylpropanoid
-
flask
- ammonia
-
prenyl
-
contigs
- pyrophosphate
- isobutyrylshikonin
The taxonomic range for the selected organisms is: Lithospermum erythrorhizon
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
lepgt1, p-hydroxybenzoate-m-geranyltransferase, p-hydroxybenzoate:geranyltransferase, aepgt6, phb geranyltransferase, lepgt-1, lepgt, aepgt, p-hydroxybenzoate geranyltransferase, diphosphate:4-hydroxybenzoate geranyltransferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
geranyl diphosphate:4-hydroxybenzoate 3-geranyltransferase
-
PHB geranyltransferase
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
geranyl-diphosphate:4-hydroxybenzoate 3-geranyltransferase
The enzyme is involved in shikonin biosynthesis. It has a strict substrate specificity for geranyl diphosphate and an absolute requirement for Mg2+ [2].
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
the enzyme accepts only geranyl diphosphate and not dimethylallyl diphosphate, farnesyl diphosphate, or geranylgeranyl diphosphate as the substrate
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
regulatory enzyme for the biosynthesis of shikonin, a naphthoquinone pigment
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
regulatory enzyme in the biosynthesis of shikonin
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
LePGT1 can utilize only geranyl diphosphate as its prenyl substrate
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
the enzyme accepts only geranyl-diphosphate and not dimethylallyl diphosphate, farnesyl diphosphate, or geranylgeranyl diphosphate as the substrate
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
the enzyme is highly specific for geranyl diphosphate and 4-hydroxybenzoate
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
LePGT1 reveals high specificity for PHB to give 3-geranyl-4-hydroxybenzoic acid (100%)
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
regulatory enzyme for the biosynthesis of shikonin, a naphthoquinone pigment
-
-
?
geranyl diphosphate + 4-hydroxybenzoate
3-geranyl-4-hydroxybenzoate + diphosphate
-
regulatory enzyme in the biosynthesis of shikonin
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
additional information
no activity with Ca2+, Cu2+, and Zn2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4-chloro-mercuriphenylsulfonic acid
-
0.1 mM, completely inhibits 4HB geranyltransferase activity
iodoacetamide
-
10 mM, completely inhibits 4HB geranyltransferase activity
N-Methylmaleimide
-
10 mM, completely inhibits 4HB geranyltransferase activity
additional information
-
blue light effectively represses the PHB geranyltransferase activity
-
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
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
Michaelis-Menten kinetics
-
0.0184
4-hydroxybenzoate
recombinant enzyme, pH 7.0-7.5, 45°C
0.0022
geranyl diphosphate
pH 7.6, 30°C, mutant enzyme D208E
0.0138
geranyl diphosphate
recombinant enzyme, pH 7.0-7.5, 45°C
0.0365
geranyl diphosphate
pH 7.6, 30°C, mutant enzyme D212E
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0665
recombinant enzyme in crude Sf9 cell extract, pH 7.0-7.5, 45°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.2 - 9.3
-
half-maximal activity at pH 6.2 and at pH 9.3, Tris-HCl buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
the mRNA of the LePGT-2 gene is undetectable in aerial plant tissues, and is exclusively detected in root tissues, similar to shikonin accumulation
the mRNA of the LePGT-1 gene is undetectable in aerial plant tissues, and is exclusively detected in root tissues, similar to shikonin accumulation
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
the reaction catalyzed by lePGT-1 plays as a competing pathway on two substrates of the Q8 synthesis pathway, namely at IPP and 4-hydroxybenzoate
physiological function
additional information
LePGT1 is a model membrane-bound aromatic substrate prenyltransferase
physiological function
-
regulatory enzyme in the biosynthesis of shikonin
physiological function
the enzyme is involved in the biosynthesis of a red naphthoquinone pigment, shikonin
physiological function
the enzyme is part of a metabolic strategy to further controll the respiratory levels in CoQ8 biosynthesis-deficient Escherichia coli strains, this metabolic strategy can lead to fine-tuning of aerobic lactate accumulation. The lactate and acetate concentrations increase with increasing enzyme levels, as does the cell density, while glucose concentration decreases, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PGT2_LITER
306
6
34222
Swiss-Prot
other Location (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
the enzyme contains contains multiple transmembrane alpha-helices
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D212A
D87A
additional information
D212A
site-directed mutagenesis, the mutant shows almost no expression of the recombinant protein
D87A
site-directed mutagenesis, the mutant can be be successfully expressed, but is catalytically inactive
additional information
the amino acid residues of LePGT1 critical for the enzymatic activity and the region responsible for the binding of the substrates are elucidated by mutational analysis. Substrate specificity is analysed using chimeric enzymes derived from LePGT1 and UbiA (EC 2.5.1.39). In vitro and in vivo analysis of the chimeras suggests that the determinant region for this specificity is within 130 amino acids of the N-terminal
additional information
the substitution of aspartate with alanine, especially D87 and D212, causes a dramatic decrease in enzyme activity
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5
-
storage at pH 5 or pH 7 results in a loss of activity between 20% and 30% in 8 days
706366
6
-
at pH 6, the enzyme can be stored in the presence of 1.5 mM digitonin, 10% glycerol and 2 mM DTT at 4°C for two weeks with minimal loss of activity
706366
7
-
storage at pH 5 or pH 7 results in a loss of activity between 20% and 30% in 8 days
706366
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, in the presence of 10% glycerol, frozen enzyme preparations are stable upon storage for at least two months. Protease inhibitors directed against serin-, carboxyl-, and metalloproteases such as PMSF, 1,2-epoxy-3-(4-nitrophenoxy)-propane or EGTA do not increase the stability of the enzyme solution.
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Of various detergents examined, digitonin is the most suitable for the solubilization of the enzyme
-
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
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
functional expression of LePGTs in a yeast COQ2 disruptant
all point mutants and chimeric enzymes are constitutively expressed in Saccharomyces cerevisiae containing a disrupted copy of the COQ2 gene
functional expression of LePGTs in a yeast COQ2 disruptant
recombinant expression in Escherichia coli coenzyme Q8 biosynthesis-deficient strains, HW108 and HW109, which contain mutations in ubiE and ubiG, respectively. Gene lePGT-1 gene is amplified and cloned into the expression vector, pBAD33, to form pBlePGT, the fine-tuning of the expression of lePGT-1 in the ubiquinone-deficient strains, HW108 and HW109, to control the electron transfer chain is investigated by adding different concentrations of the inducer arabinose at 0-20 mM under fully aerobic conditions
recombinant expression of C-terminally His-tagged LePGT1 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection method
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Gaisser, S.; Heide, L.
Inhibition and regulation of shikonin biosynthesis in suspension cultures of Lithospermum
Phytochemistry
41
1065-1072
1996
Lithospermum erythrorhizon
-
Ohara, K.; Muroya, A.; Fukushima, N.; Yazaki, K.
Functional characterization of LePGT1, a membrane-bound prenyltransferase involved in the geranylation of p-hydroxybenzoic acid
Biochem. J.
421
231-241
2009
Lithospermum erythrorhizon (Q8W405)
Yazaki, K.; Kunihisa, M.; Fujisaki, T.; Sato, F.
Geranyl diphosphate:4-hydroxybenzoate geranyltransferase from Lithospermum erythrorhizon. Cloning and characterization of a ket enzyme in shikonin biosynthesis
J. Biol. Chem.
277
6240-6246
2001
Lithospermum erythrorhizon (Q8W404), Lithospermum erythrorhizon (Q8W405), Lithospermum erythrorhizon
Yamaga, Y.; Nakanishi, K.; Fukui, H.; Tabata, M.
Intracellular localization of p-hydroxybenzoate geranyltransferase, a key enzyme involved in shikonin biosynthesis
Phytochemistry
32
633-636
1993
Lithospermum erythrorhizon
-
Mhlenweg, A.; Melzer, M.; Li, S.M.; Heide, L.
4-Hydroxybenzoate 3-geranyltransferase from Lithospermum erythrorhizon: purification of a plant membrane-bound prenyltransferase
Planta
205
407-413
1998
Lithospermum erythrorhizon
Wu, H.; Bennett, G.N.; San, K.Y.
Metabolic control of respiratory levels in coenzyme Q biosynthesis-deficient Escherichia coli strains leading to fine-tune aerobic lactate fermentation
Biotechnol. Bioeng.
112
1720-1726
2015
Lithospermum erythrorhizon (Q8W405), Lithospermum erythrorhizon
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
FEBS J.
280
2572-2580
2013
Lithospermum erythrorhizon (Q8W405)
-
EXTERNAL LINKS (specific for EC-Number 2.5.1.93)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
