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(2E,6E)-8-O-(N-methyl-2-aminobenzoyl)-3,7-dimethyl-2,6-octandien-1-diphosphate + 5 isopentenyl diphosphate
?
-
-
-
?
(2E,6E)-farnesyl diphosphate + 3-bromo-3-butenyl diphosphate
diphosphate + ?
-
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
(2E,6E)-farnesyl diphosphate + 6 isopentenyl diphosphate
6 diphosphate + all-trans-nonaprenyl diphosphate
-
-
-
?
dimethylallyl diphosphate + isopentenyl diphosphate
diphosphate + octaprenyl diphosphate + nonaprenyl diphosphate
geranyl diphosphate + isopentenyl diphosphate
?
geranylgeranyl diphosphate + isopentenyl diphosphate
?
additional information
?
-
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
80% all-trans-octaprenyl diphosphate, 20% nonaprenyl diphosphate in presence of 0.01 mM enzyme, 0.0001 mM farnesyl diphosphate, 0.001 mM isopentenyl diphosphate. Variation of concentrations and conditions allows for synthesis of products with chain length C20 to C60
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
the enzyme catalyzes the successive condensation of farnesyl diphosphate with five isopentenyl pyrophosphate molecules to form trans-C40-octaprenyl diphosphate
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
(2E,6E)-farnesyl diphosphate is the most effective substrate
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
(2E,6E)-farnesyl diphosphate is the most effective substrate
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
-
-
the product of chain elongation is C40 and the rate of its conversion to C45 is negligible. Under single-turnover condition with 0.01 mM enzyme-farnesyl diphosphate complex and 0.001 mM isopentenyl diphosphate, only the C20 is formed
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
the polymer all-trans C40-octaprenyl pyrophosphate forms the side chain of ubiquinone that is involved in electron transport system to produce ATP
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
the product of chain elongation catalyzed by the Thermotoga maritima enzyme is C40 and the rate of its conversion to C45 is negligible. Under single-turnover condition with 0.01 mM octaprenyl pyrophosphate synthase/farnesyl pyrophosphate complex and 0.001 mM isopentenyl pyrophosphate, only the C20 is formed
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
the polymer all-trans C40-octaprenyl pyrophosphate forms the side chain of ubiquinone that is involved in electron transport system to produce ATP
-
-
?
(2E,6E)-farnesyl diphosphate + 5 isopentenyl diphosphate
5 diphosphate + all-trans-octaprenyl diphosphate
the product of chain elongation catalyzed by the Thermotoga maritima enzyme is C40 and the rate of its conversion to C45 is negligible. Under single-turnover condition with 0.01 mM octaprenyl pyrophosphate synthase/farnesyl pyrophosphate complex and 0.001 mM isopentenyl pyrophosphate, only the C20 is formed
-
-
?
dimethylallyl diphosphate + isopentenyl diphosphate
diphosphate + octaprenyl diphosphate + nonaprenyl diphosphate
0.7% activity compared to (2E,6E)-farnesyl diphosphate
-
-
?
dimethylallyl diphosphate + isopentenyl diphosphate
diphosphate + octaprenyl diphosphate + nonaprenyl diphosphate
0.7% activity compared to (2E,6E)-farnesyl diphosphate
-
-
?
geranyl diphosphate + isopentenyl diphosphate
?
5.3% activity compared to (2E,6E)-farnesyl diphosphate
-
-
?
geranyl diphosphate + isopentenyl diphosphate
?
5.3% activity compared to (2E,6E)-farnesyl diphosphate
-
-
?
geranylgeranyl diphosphate + isopentenyl diphosphate
?
53.4% activity compared to (2E,6E)-farnesyl diphosphate
-
-
?
geranylgeranyl diphosphate + isopentenyl diphosphate
?
53.4% activity compared to (2E,6E)-farnesyl diphosphate
-
-
?
additional information
?
-
at the bottom of the active-site tunnel, residues M123 and M135 act in concert to form a wall which determines the final chain length, substrate binding and enzyme-ligand structure analysis, overview
-
-
?
additional information
?
-
-
at the bottom of the active-site tunnel, residues M123 and M135 act in concert to form a wall which determines the final chain length, substrate binding and enzyme-ligand structure analysis, overview
-
-
?
additional information
?
-
no substrate: dimethylallyl diphosphate, geranyl diphosphate
-
-
?
additional information
?
-
residues Tyr-38, Lys-235, and Arg-321 are involved in chain length determination
-
-
?
additional information
?
-
no substrate: geranyl diphosphate, geranylgeranyl diphosphate
-
-
?
additional information
?
-
-
no substrate: geranyl diphosphate, geranylgeranyl diphosphate
-
-
?
additional information
?
-
-
the isopentenyl diphosphate condensation represents the rate-limiting step of the reaction
-
-
?
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0.0008
(2E,6E)-8-O-(N-methyl-2-aminobenzoyl)-3,7-dimethyl-2,6-octandien-1-diphosphate
at pH 7.5, temperature not specified in the publication
0.00025 - 0.0246
(2E,6E)-farnesyl diphosphate
0.0007
dimethylallyl diphosphate
at pH 7.5 and 30°C
0.00018 - 0.34
isopentenyl diphosphate
0.00025
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C
0.0008
(2E,6E)-farnesyl diphosphate
mutant K225L, pH 7.5, 25°C
0.0008
(2E,6E)-farnesyl diphosphate
mutant Q208A, pH 7.5, 25°C
0.001
(2E,6E)-farnesyl diphosphate
mutant K45A, pH 7.5, 25°C
0.0011
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C
0.0012
(2E,6E)-farnesyl diphosphate
mutant T171V, pH 7.5, 25°C
0.0015
(2E,6E)-farnesyl diphosphate
-
pH 7.5, 25°C
0.0015
(2E,6E)-farnesyl diphosphate
pH 7.5, 25°C
0.0015
(2E,6E)-farnesyl diphosphate
wild-type, pH 7.5, 25°C
0.0015
(2E,6E)-farnesyl diphosphate
mutant D215A, pH 7.5, 25°C
0.0015
(2E,6E)-farnesyl diphosphate
at pH 7.5, temperature not specified in the publication
0.0017
(2E,6E)-farnesyl diphosphate
mutant D211A, pH 7.5, 25°C
0.0018
(2E,6E)-farnesyl diphosphate
mutant K235L, pH 7.5, 25°C
0.0028
(2E,6E)-farnesyl diphosphate
mutant R94A, pH 7.5, 25°C
0.0031
(2E,6E)-farnesyl diphosphate
mutant D212A, pH 7.5, 25°C
0.0032
(2E,6E)-farnesyl diphosphate
mutant K170L, pH 7.5, 25°C
0.0038
(2E,6E)-farnesyl diphosphate
mutant D88A, pH 7.5, 25°C
0.0052
(2E,6E)-farnesyl diphosphate
mutant R93A, pH 7.5, 25°C
0.0092
(2E,6E)-farnesyl diphosphate
mutant D84A, pH 7.5, 25°C
0.024
(2E,6E)-farnesyl diphosphate
mutant D85A, pH 7.5, 25°C
0.0242
(2E,6E)-farnesyl diphosphate
mutant H77A, pH 7.5, 25°C
0.0246
(2E,6E)-farnesyl diphosphate
mutant R48A, pH 7.5, 25°C
0.00018
isopentenyl diphosphate
pH 7.5, 30°C
0.002
isopentenyl diphosphate
-
pH 7.5, 25°C
0.002
isopentenyl diphosphate
pH 7.5, 25°C
0.0024
isopentenyl diphosphate
pH 7.5, 30°C
0.0024
isopentenyl diphosphate
mutant Q208A, pH 7.5, 25°C
0.0037
isopentenyl diphosphate
mutant R93A, pH 7.5, 25°C
0.004
isopentenyl diphosphate
wild-type, pH 7.5, 25°C
0.004
isopentenyl diphosphate
with (2R,6E)-farnesyl diphosphate as cosubstrate, at pH 7.5, temperature not specified in the publication
0.0045
isopentenyl diphosphate
mutant T171V, pH 7.5, 25°C
0.0049
isopentenyl diphosphate
mutant K170L, pH 7.5, 25°C
0.0069
isopentenyl diphosphate
with (2E,6E)-8-O-(N-methyl-2-aminobenzoyl)-3,7-dimethyl-2,6-octandien-1-diphosphate as cosubstrate, at pH 7.5, temperature not specified in the publication
0.0085
isopentenyl diphosphate
mutant D88A, pH 7.5, 25°C
0.0093
isopentenyl diphosphate
mutant D84A, pH 7.5, 25°C
0.0113
isopentenyl diphosphate
mutant K225L, pH 7.5, 25°C
0.0182
isopentenyl diphosphate
mutant D215A, pH 7.5, 25°C
0.0233
isopentenyl diphosphate
mutant D212A, pH 7.5, 25°C
0.0271
isopentenyl diphosphate
mutant D211A, pH 7.5, 25°C
0.029
isopentenyl diphosphate
mutant D85A, pH 7.5, 25°C
0.0335
isopentenyl diphosphate
mutant K45A, pH 7.5, 25°C
0.0539
isopentenyl diphosphate
mutant R94A, pH 7.5, 25°C
0.0639
isopentenyl diphosphate
mutant R48A, pH 7.5, 25°C
0.0745
isopentenyl diphosphate
mutant K235L, pH 7.5, 25°C
0.102
isopentenyl diphosphate
mutant H77A, pH 7.5, 25°C
0.34
isopentenyl diphosphate
with dimethylallyl diphosphate as cosubstrate, at pH 7.5 and 30°C
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0.055
(2E,6E)-8-O-(N-methyl-2-aminobenzoyl)-3,7-dimethyl-2,6-octandien-1-diphosphate
at pH 7.5, temperature not specified in the publication
0.00000023 - 2
(2E,6E)-farnesyl diphosphate
0.005
isopentenyl diphosphate
pH 7.5, 25°C
0.00000023
(2E,6E)-farnesyl diphosphate
mutant A76Y/S77F, pH 7.5, 25°C
0.00000029
(2E,6E)-farnesyl diphosphate
mutant S77F, pH 7.5, 25°C
0.000073
(2E,6E)-farnesyl diphosphate
mutant A76Y, pH 7.5, 25°C
0.000084
(2E,6E)-farnesyl diphosphate
mutant R94A, pH 7.5, 25°C
0.00011
(2E,6E)-farnesyl diphosphate
mutant D88A, pH 7.5, 25°C
0.00012
(2E,6E)-farnesyl diphosphate
mutant D84A, pH 7.5, 25°C
0.00012
(2E,6E)-farnesyl diphosphate
mutant R93A, pH 7.5, 25°C
0.00032
(2E,6E)-farnesyl diphosphate
mutant D85A, pH 7.5, 25°C
0.00033
(2E,6E)-farnesyl diphosphate
mutant K170L, pH 7.5, 25°C
0.0011
(2E,6E)-farnesyl diphosphate
mutant R48A, pH 7.5, 25°C
0.0018
(2E,6E)-farnesyl diphosphate
mutant D212A, pH 7.5, 25°C
0.0019
(2E,6E)-farnesyl diphosphate
mutant K235L, pH 7.5, 25°C
0.0033
(2E,6E)-farnesyl diphosphate
mutant V37Y, pH 7.5, 25°C
0.0035
(2E,6E)-farnesyl diphosphate
mutant F52A, pH 7.5, 25°C
0.0036
(2E,6E)-farnesyl diphosphate
mutant K45A, pH 7.5, 25°C
0.0043
(2E,6E)-farnesyl diphosphate
mutant T171V, pH 7.5, 25°C
0.005
(2E,6E)-farnesyl diphosphate
-
pH 7.5, 25°C
0.005
(2E,6E)-farnesyl diphosphate
pH 7.5, 25°C
0.005
(2E,6E)-farnesyl diphosphate
wild-type, pH 7.5, 25°C
0.0051
(2E,6E)-farnesyl diphosphate
mutant F132A, pH 7.5, 25°C
0.0075
(2E,6E)-farnesyl diphosphate
mutant D211A, pH 7.5, 25°C
0.0087
(2E,6E)-farnesyl diphosphate
mutant D215A, pH 7.5, 25°C
0.0091
(2E,6E)-farnesyl diphosphate
mutant K225L, pH 7.5, 25°C
0.0096
(2E,6E)-farnesyl diphosphate
mutant Q208A, pH 7.5, 25°C
0.019
(2E,6E)-farnesyl diphosphate
mutant H77A, pH 7.5, 25°C
0.06
(2E,6E)-farnesyl diphosphate
at pH 7.5, temperature not specified in the publication
2
(2E,6E)-farnesyl diphosphate
wild-type, pH 7.5, 25°C
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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
evolution
octaprenyl diphosphate synthase, OPP, belongs to the trans-prenyltransferase class of protein
evolution
the enzyme belongs to the E-type prenyltransferase family
metabolism
octaprenyl pyrophosphate synthase catalyzes the chain elongation of farnesyl pyrophosphate via consecutive condensation reactions with five molecules of isopentenyl pyrophosphate to generate all-trans C40-octaprenyl pyrophosphate. The polymer forms the side chain of ubiquinone that is involved in electron transport system to produce ATP
metabolism
-
octaprenyl pyrophosphate synthase catalyzes the chain elongation of farnesyl pyrophosphate via consecutive condensation reactions with five molecules of isopentenyl pyrophosphate to generate all-trans C40-octaprenyl pyrophosphate. The polymer forms the side chain of ubiquinone that is involved in electron transport system to produce ATP
-
physiological function
-
octaprenyl diphosphate synthase is responsible for the synthesis of the side chain of isoprenoid quinones
physiological function
the enzyme catalyzes consecutive condensation reactions of farnesyl diphosphate with isopentenyl diphosphate to generate octaprenyl diphosphate, which constitutes the side chain of bacterial ubiquinone or menaquinone
physiological function
the chromosomal ispB disrupted mutant cannot be isolated unless the ispB gene or its homolog is supplied on a plasmid. The chromosomal ispB disruptants that harbor plasmids carrying the ispB homologs from Haemophilus influenzae and Synechocystis sp. strain PCC6803 produce mainly ubiquinone 7 and ubiquinone 9, respectively
physiological function
gene product of gbs1789 is involved in the biosynthesis of long-chain demethylmenaquinones. A gbs1789 mutant cannot respire in the presence of heme and dihydroxy-2-naphthoic acid. Isoprenoid side chains from Group B Streptococcus demethylmenaquinones are produced by the protein encoded by the gbs1783 gene, since this gene can complement an Escherichia coli ispB mutant defective for isoprenoids chain synthesis
physiological function
the enzyme catalyzes consecutive condensation reactions of one allylic substrate farnesyl diphosphate and five homoallylic substrate isopentenyl diphosphate molecules to form a C40 long-chain product octaprenyl diphosphate, which serves as a side chain of ubiquinone and menaquinone
physiological function
the enzyme is essential for the biosynthesis of bacterial ubiquinone or menaquinone side chains, which play an important role in the electron-transport system
physiological function
Streptococcus agalactiae serogroup III NEM316
-
gene product of gbs1789 is involved in the biosynthesis of long-chain demethylmenaquinones. A gbs1789 mutant cannot respire in the presence of heme and dihydroxy-2-naphthoic acid. Isoprenoid side chains from Group B Streptococcus demethylmenaquinones are produced by the protein encoded by the gbs1783 gene, since this gene can complement an Escherichia coli ispB mutant defective for isoprenoids chain synthesis
-
additional information
enzyme homology modeling, overview. (2E,6E)-Farnesyl diphosphate is bound to the first DDXXD motif, and isopentenyl diphosphate is bound to several cationic residues (e.g. Arg, Lys, and His) via diphosphate, but not to the second DDXXD motif. The enzyme contains an elongated tunnel-shaped crevice as the active site to accommodate its reactants and product
additional information
-
enzyme homology modeling, overview. (2E,6E)-Farnesyl diphosphate is bound to the first DDXXD motif, and isopentenyl diphosphate is bound to several cationic residues (e.g. Arg, Lys, and His) via diphosphate, but not to the second DDXXD motif. The enzyme contains an elongated tunnel-shaped crevice as the active site to accommodate its reactants and product
additional information
enzyme structure modelling using the structure of Rhodobacter capsulatus decaprenyl diphosphate synthase, PDB ID 3mzv, as a template model
additional information
-
enzyme structure modelling using the structure of Rhodobacter capsulatus decaprenyl diphosphate synthase, PDB ID 3mzv, as a template model
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purified recombinant enzyme, crystallization method screening, sitting drop vapour diffusion method, mixing of 0.002 ml of 3 mg/ml protein in 25 mM Tris-HCl, 150 mM NaCl, pH 7.5, with 0.002 ml of crystallization solution containing 0.3 M magnesium chloride hexahydrate, 0.1 M Tris-HCl, pH 8.5, 24% w/v PEG 3350, 3-4 days, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement, the crystal contains one homodimer per asymmetric unit, structure modelling
purified recombinant His-tagged enzyme in apo-form and in complexes with isopentenyl diphosphate and a farnesyl diphosphate thio-analogue, FsPP, sitting drop vapor diffusion method, mixing of 0.002 ml of 3 mg/ml protein in 25 mM Tris-HCl, pH 7.5, and 150 mM NaCl, with 0.002 ml of reservoir solution containing 0.3 M magnesium chloride, 0.1 M Tris-HCl, pH 8.5, and 24% w/v PEG 3350, and equilibration against 0.3 ml of reservori solution, 22°C, 3-4 days, X-ray diffraction structure determination and analysis at resolutions of 2.2-2.6 A
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
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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 30°C, no growth at 43°C, main product is ubiquinone-8
R321D
normal growth at 30°C, no growth at 43°C
R321V
products are ubiquinone-6 and ubiquinone-7, slow growth at 43°C
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
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
V73Y
mutation leads to additional accumulation of C30 intermediate
A76Y
main product is C20
A76Y
mutant produces only C20 cores instead of the C40 core of octaprenyl diphosphate
A76Y/S77F
main product is C20
A76Y/S77F
mutant produces only C20 cores instead of the C40 core of octaprenyl diphosphate. The A76Y/S77F mutant synthesizes a larger amount of C20 than the A76Y mutant
F132A
product is predominantly C50
F132A
steady-state activity 0.0051 per s. Product chain length C45 to C60
additional information
for biotechnological production of coenzyme Q10 in recombinant Escherichia coli, three genetic manipulations are performed: heterologous expression of decaprenyl diphosphate synthase (Dps) from Agrobacterium tumefaciens, deletion of endogenous octaprenyl diphosphate synthase (IspB), and overexpression of 1-deoxy-D-xylulose synthase (Dxs). Expression of the dps gene and deletion of the ispB gene in Escherichia coli BL21(DE3)DELTAispB/pAP1 allows production of CoQ10 only. Coexpression of the dxs gene increases the specific content of CoQ10
additional information
-
for biotechnological production of coenzyme Q10 in recombinant Escherichia coli, three genetic manipulations are performed: heterologous expression of decaprenyl diphosphate synthase (Dps) from Agrobacterium tumefaciens, deletion of endogenous octaprenyl diphosphate synthase (IspB), and overexpression of 1-deoxy-D-xylulose synthase (Dxs). Expression of the dps gene and deletion of the ispB gene in Escherichia coli BL21(DE3)DELTAispB/pAP1 allows production of CoQ10 only. Coexpression of the dxs gene increases the specific content of CoQ10
additional information
residue D85 is the most important residue in the first DDXXD motif for both farnesyl diphosphate and isopentenyl diphosphate binding through an H-bond network involving R93 and R94, respectively, whereas R94, K45, R48, and H77 are responsible for isopentenyl binding by providing H-bonds and ionic interactions. K170 and T171 may stabilize the farnesyl carbocation intermediate to facilitate the reaction, whereas R93 and K225 may stabilize the catalytic base for HR proton abstraction after isopentenyl diphosphate condensation. K225 and K235 in a flexible loop may interact with farnesyl diphosphate when the enzyme becomes a closed conformation, which is therefore crucial for catalysis. Q208 is near the hydrophobic part of isopentenyl diphosphate and is important for isopentenyl diphosphate binding and catalysis
additional information
-
residue D85 is the most important residue in the first DDXXD motif for both farnesyl diphosphate and isopentenyl diphosphate binding through an H-bond network involving R93 and R94, respectively, whereas R94, K45, R48, and H77 are responsible for isopentenyl binding by providing H-bonds and ionic interactions. K170 and T171 may stabilize the farnesyl carbocation intermediate to facilitate the reaction, whereas R93 and K225 may stabilize the catalytic base for HR proton abstraction after isopentenyl diphosphate condensation. K225 and K235 in a flexible loop may interact with farnesyl diphosphate when the enzyme becomes a closed conformation, which is therefore crucial for catalysis. Q208 is near the hydrophobic part of isopentenyl diphosphate and is important for isopentenyl diphosphate binding and catalysis
additional information
expression rescues an Escherichia coli chromosomal ispB disruptant
additional information
-
expression rescues an Escherichia coli chromosomal ispB disruptant
additional information
-
expression rescues an Escherichia coli chromosomal ispB disruptant
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