Information on EC 2.5.1.90 - all-trans-octaprenyl-diphosphate synthase

Word Map on EC 2.5.1.90
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
2.5.1.90
-
RECOMMENDED NAME
GeneOntology No.
all-trans-octaprenyl-diphosphate synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate = 5 diphosphate + all-trans-octaprenyl diphosphate
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
octaprenyl diphosphate biosynthesis
-
-
ubiquinone biosynthesis
-
-
Terpenoid backbone biosynthesis
-
-
Biosynthesis of secondary metabolites
-
-
SYSTEMATIC NAME
IUBMB Comments
(2E,6E)-farnesyl-diphosphate:isopentenyl-diphosphate farnesyltranstransferase (adding 5 isopentenyl units)
This enzyme catalyses the condensation reactions resulting in the formation of all-trans-octaprenyl diphosphate, the isoprenoid side chain of ubiquinone-8 and menaquinone-8. The enzyme adds five isopentenyl diphosphate molecules sequentially to farnesyl diphosphate with trans stereochemistry
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
it is impossible to obtain an ispB deletion mutant unless the ispB gene or its homolog is supplied on a plasmid. The ispB gene is essential for the normal growth of Escherichia coli
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2E,6E)-farnesyl diphosphate + 3-bromo-3-butenyl diphosphate
diphosphate + ?
show the reaction diagram
-
isopentenyl diphosphate analogue, showing a significantly reduced activity. Only two 3-bromo-3-butenyl diphosphate condensation reactions occurr in 24 h and the reaction products do not reach C40 and C55. Use results in trapping of farnesol in the reaction from radioloabeled farnesyl diphosphate under basic conditions, consistent with a sequential mechanism
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
show the reaction diagram
(2E,6E)-farnesyl diphosphate + 6 isopentenyl diphosphate
6 diphosphate + all-trans-nonaprenyl diphosphate
show the reaction diagram
-
-
-
?
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
nerolidol
competitive, 57% inhibition at 50 nM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Triton X-100
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00025 - 0.0246
(2E,6E)-farnesyl diphosphate
0.00018 - 0.102
isopentenyl diphosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000023 - 2
(2E,6E)-farnesyl diphosphate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000015
nerolidol
pH 7.5, 30C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 63000, SDS-PAGE of recombinant enzyme
additional information
-
IspB interacts with Schizosaccharomyces pombe decaprenyl diphosphate synthase Dps1 and D-less polyprenyl diphosphate synthase Dlp1, forming a high-molecular weight complex that stabilizes IspB
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme and selenomethionine derivative, to 2.0 A and 2.8 A resolution, respectively. Residues Arg87, Lys36 and Arg39 are essential for isopentenyl diphosphate binding. Residues Lys162, Lys224 and Gln197 are involved in farnesyl diphosphate binding. The second DDXXD motif may be involved in farnesyl diphosphate binding by Mg2+-mediated interactions, Leu127 is probably involved in product chain length determination and the intermediate products such as geranylgeranyl diphosphate need a rearrange to occupy the binding site of farnesyl diphosphate and then isopentenyl diphosphate is reloaded
mutants A76Y, A76Y/S77F, F132A/L128A, F132A/L128A/I123A, and F132A/L128A/I123A/D62A to 3.1, 2.7, 3.3, 3.35 and 3.4 A resolution, respectively. Like wildtype OPPs, all mutant structures contain 12 alpha-helices, nine of them surrounding a large central cavity and an elongated tunnel-shaped active site cavity surrounded by four alpha-helices In the crystal structure of the A76Y/S77F mutant, F77 is pushed away by Y76, thereby creating more space between those two large amino acids to accommodate the C20 product. A large F132 residue at the bottom of the tunnel-shaped active site serves as the floor and determines the final product chain length. The substitution of F132 with a small Ala, thereby removing the blockade, leads to the synthesis of a C50 product larger than that produced by the wild-type enzyme
-
wild-type and mutants F52A, V73A, S77F, F132A, to 2.28, 2.80, 2.85, 2.45 and 2.40 A resolution, respectively. OPPs is composed entirely of alpha-helices joined by connecting loops and is arranged with nine core helices around a large central cavity. An elongated hydrophobic tunnel between D and F alpha-helices contains two DDXXD motifs on the top for substrate binding and is occupied at the bottom with two large residues Phe-52 and Phe-132
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
-
melting temeprature is abve 80C
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partial, from schizont stage parasite
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression as glutathione-S-transferase fusion protein
expression in Escherichia coli
-
expression in Escherichia coli and Saccharomyces cerevisiae
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D211A
-
0.37% of wild-type activity
D212A
-
0.091% of wild-type activity
D215A
-
0.43% of wild-type activity
D84A
-
0.006% of wild-type activity
D85A
-
0.016% of wild-type activity
D88A
-
0.0057% of wild-type activity
H77A
-
0.95% of wild-type activity
K170L
-
0.017% of wild-type activity
K225L
-
0.45% of wild-type activity
K235L
-
0.096% of wild-type activity
K45A
-
0.18% of wild-type activity
Q208A
-
0.048% of wild-type activity
R48A
-
0.057% of wild-type activity
R93A
-
0.0058% of wild-type activity
R94A
-
0.0042% of wild-type activity
T171V
-
0.22% of wild-type activity
A79Y
-
no activity. Cells harboring wild-type ispB and the A79Y mutant produce mainly ubiquinone-6, although the activity of the enzyme with the A79Y mutation is completely abolished. Although the A79Y mutant is functionally inactive, it can regulate activity upon forming a heterodimer with wild-type IspB, and this dimer formation is important for the determination of the isoprenoid chain length
F75A
-
resulting ubiquinone species are almost the same as those produced by the wild-type enzyme
I32V
-
resulting ubiquinone species are almost the same as those produced by the wild-type enzyme
K170A
-
mutant cannot be isolated, mutant protein does not retain functional activity
K170G
-
mutant cannot be isolated, mutant protein does not retain functional activity
K235L
-
main product is ubiquinone-8
L31V
-
resulting ubiquinone species are almost the same as those produced by the wild-type enzyme
R321A
-
normal growth at 30C, no growth at 43C, main product is ubiquinone-8
R321D
-
normal growth at 30C, no growth at 43C
R321V
-
products are ubiquinone-6 and ubiquinone-7, slow growth at 43C
Y37A
-
mutant cannot be isolated, mutant protein does not retain functional activity
Y37A/Y38A
-
increased production of ubiquinone-6
Y38A
-
products are ubiquinone-6 and ubiquinone-7
Y38A/R321V
-
products are ubiquinone-5 and ubiquinone-6
Y61V
-
resulting ubiquinone species are almost the same as those produced by the wild-type enzyme
R321A
-
expression of Schizosaccharomyces pombe decaprenyl diphosphate synthase Dps1 or D-less polyprenyl diphosphate synthase Dlp1 recover the thermo-sensitive growth of an Escherichia coli ispB R321A mutant and restor IspB activity and production of coenzyme Q-8. IspB interacts with Dlp1 or Dps1, forming a high-molecular weight complex that stabilizes IspB, leading to full functionality
A76Y/S77F
F132A/L128A
-
steady-state activity 0.0008 per s. Product chain length C55, C60
F132A/L128A/I123A
-
steady-state activity 0.00066 per s. Produuct chain length C55 to C75
F132A/L128A/I123A/D62A
-
products reach C95, beyond the largest chain length generated by all known trans-prenyltransferases. Steady-state activity 0.00061 per s
F52A
-
product is predominatly C40, like in wild-type
S77F
-
main product is C20
V73Y
-
mutation leads to additional accumulation of C30 intermediate
additional information
Show AA Sequence (724 entries)
Please use the Sequence Search for a certain query.