Information on EC 2.5.1.30 - heptaprenyl diphosphate synthase

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The expected taxonomic range for this enzyme is: Bacillaceae

EC NUMBER
COMMENTARY
2.5.1.30
-
RECOMMENDED NAME
GeneOntology No.
heptaprenyl diphosphate synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate = 4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
-
-
-
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate = 4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
enzyme component I, enzyme component II, and farnesyl diphosphate-Mg2+ form a ternary complex during catalysis and that neither isopentenyl diphosphate nor the product, heptaprenyl diphosphate, is included in this complex, which probably represents a catalytically active state of the HepPP synthase. Component I is involved in allylic substrate recognition
-
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate = 4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
initially the substrate farnesyl diphosphate binds to subunit II using Mg2+, followed by the formation of the subunit I-farnesyl diphosphate-Mg2+-subunit II complex
-
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate = 4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
the C-C bond formation in the heptaprenyl diphosphate synthase reaction takes place at the si face of the double bond of isopentenyl diphosphate
-
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate = 4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
a common mechanism controls the product chain length of both short-chain and medium-chain prenyl diphosphate synthases. In wild-type heptaprenyl diphosphate synthase, the prenyl chain can grow on the surface of the small residues at positions 79 and 80, and the elongation is precisely blocked at the length of C35 by Ile76
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
heptaprenyl diphosphate biosynthesis
-
Terpenoid backbone biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
(2E,6E)-farnesyl-diphosphate:isopentenyl-diphosphate farnesyltranstransferase (adding 4 isopentenyl units)
This enzyme catalyses the condensation reactions resulting in the formation of all-trans-heptaprenyl diphosphate, the isoprenoid side chain of ubiquinone-7 and menaquinone-7. The enzyme adds four isopentenyl diphosphate molecules sequentially to farnesyl diphosphate with trans stereochemistry.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
HepPP synthase
P31112 and P31114
-
HepPP synthase
P55784 and P55785
-
HepPS
P31112 and P31114
-
heptaprenyl pyrophosphate synthetase
P31112 and P31114
-
heptaprenylpyrophosphate synthetase
P31112 and P31114
-
heptaprenylpyrophosphate synthetase
Bacillus subtilis PCI-219
P31112 and P31114
-
-
CAS REGISTRY NUMBER
COMMENTARY
74506-59-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
P31112: subunit 1 (gerC1), P31114: subunit 2 (gerC3)
P31112 and P31114
SwissProt
Manually annotated by BRENDA team
P31112: subunit 1, P31114: subunit 2
P31112 and P31114
SwissProt
Manually annotated by BRENDA team
P31112: subunit 1, P31114: subunit 2; PCI-219
P31112 and P31114
SwissProt
Manually annotated by BRENDA team
Bacillus subtilis PCI-219
P31112: subunit 1, P31114: subunit 2; PCI-219
P31112 and P31114
SwissProt
Manually annotated by BRENDA team
P55784: subunit 1, P55785: subunit 2
P55784 and P55785
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
P31112 and P31114
heptaprenyl diphosphate synthase is involved in the biosynthesis of the side chain of menaquinone-7
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2E,6E)-farnesyl diphosphate + 3 isopentenyl diphosphate
3 diphosphate + all-trans-hexaprenyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 3-ethylbut-3-enyl diphosphate
diphosphate + (all-E)-3-ethyl-7,11,15-trimethylhexadeca-2,6,10,14-tetraenyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 3-propylbut-3-enyl diphosphate
diphosphate + (all-E)-3-propyl-7,11,15-trimethylhexadeca-2,6,10,14-tetraenyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P31112 and P31114
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
P31112 and P31114
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P55784 and P55785
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
P31112 and P31114
heptaprenyl diphosphate synthase is involved in the biosynthesis of the side chain of menaquinone-7
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P31112 and P31114
synthesis of all-trans-heptaprenyl diphosphate, which is precursor for the side chain of menaquinone-7
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
initially the substrate farnesyl diphosphate binds to subunit II using Mg2+, followed by the formation of the subunit I-farnesyl diphosphate-Mg2+-subunit II complex
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
the C-C bond formation in the heptaprenyl diphosphate synthase reaction takes place at the si face of the double bond of isopentenyl diphosphate
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P31112 and P31114
the enzyme reaction proceeds with an elimination of 2-pro-R hydrogen of isopentenyl diposphate without accumulation of any prenyl diphosphate shorter than C35
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
wild-type enzyme mainly produces heptaprenyl diphosphate (C35, molar ratio, 73%) and does not yield any product longer than heptaprenyl diphosphate
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
Bacillus subtilis PCI-219
P31112 and P31114
-, synthesis of all-trans-heptaprenyl diphosphate, which is precursor for the side chain of menaquinone-7
-
-
?
(2E,6E)-farnesyl diphosphate + but-3-enyl diphosphate
diphosphate + (E)-norgeranylgeranyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + but-3-enyl diphosphate
diphosphate + E-norgeranylgeranyl diphosphate
show the reaction diagram
-
-
-
-
?
all-trans-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
enzyme component I, enzyme component II, and farnesyl diphosphate-Mg2+ form a ternary complex during catalysis and that neither isopentenyl diphosphate nor the product, heptaprenyl diphosphate, is included in this complex, which probably represents a catalytically active state of the HepPP synthase. Enzyme component I is involved in allylic substrate recognition
-
-
?
geranyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-hexaprenyl diphosphate
show the reaction diagram
-
geranyl diphosphate is a poor substrate
-
-
?
geranylgeranyl diphosphate + 2 isopentenyl diphosphate
2 diphosphate + all-trans-hexaprenyl diphosphate
show the reaction diagram
-
-
-
-
?
geranylgeranyl diphosphate + 3 isopentenyl diphosphate
3 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P31112 and P31114
-
-
-
?
geranylgeranyl diphosphate + 3 isopentenyl diphosphate
3 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
-
-
-
?
geranylgeranyl diphosphate + 3 isopentenyl diphosphate
3 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P31112 and P31114
the enzyme reaction proceeds with an elimination of 2-pro-R hydrogen of isopentenyl diphosphate without accumulation of any prenyl diphosphate shorter than C35. Poor activity of geranylneryl diphosphate contrast with the high activity of the all-trans isomer is consistent with the proposal that this enzyme catalyzes condensations resulting in trans products
-
-
?
geranylgeranyl diphosphate + 3 isopentenyl diphosphate
3 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
Bacillus subtilis PCI-219
P31112 and P31114
-
-
-
?
geranylgeranyl diphosphate + isopentenyl diphosphate
diphosphate + all-trans-heptaprenyl diphosphate + geranylfarnesyl diphosphate
show the reaction diagram
-
-
wild-type enzyme mainly yields geranylfarnesyl diphosphate (C25, 59%) with a significant amount of heptaprenyl diphosphate (37%)
-
?
additional information
?
-
-
dimethylallyl diphosphate and geranyl diphosphate, are almost inactive as substrates
-
-
-
additional information
?
-
-, P31112 and P31114
neither dimethylallyl diphosphate nor geranyl diphosphate is active as primer
-
-
-
additional information
?
-
-
no activity with 3-butylbut-3-enyl diphosphate, norfarnesyl diphosphate or norgeranylgeranyl diphosphate
-
-
-
additional information
?
-
-
norfarnesyl diphosphate is not accepted as substrate
-
-
-
additional information
?
-
-, P31112 and P31114
the enzyme does not catalyze a reaction between isopentenyl diphosphate and either dimethylallyl or geranyl diphosphate
-
-
-
additional information
?
-
Bacillus subtilis PCI-219
P31112 and P31114
neither dimethylallyl diphosphate nor geranyl diphosphate is active as primer
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-
-
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
P31112 and P31114
heptaprenyl diphosphate synthase is involved in the biosynthesis of the side chain of menaquinone-7
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
-, P31112 and P31114
synthesis of all-trans-heptaprenyl diphosphate, which is precursor for the side chain of menaquinone-7
-
-
?
(2E,6E)-farnesyl diphosphate + 4 isopentenyl diphosphate
4 diphosphate + all-trans-heptaprenyl diphosphate
show the reaction diagram
Bacillus subtilis PCI-219
P31112 and P31114
synthesis of all-trans-heptaprenyl diphosphate, which is precursor for the side chain of menaquinone-7
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
required, no activation with Mn2+
Mg2+
P31112 and P31114
absolute requirement
Mg2+
-
requires a divalent cation for the enzymatic activity with an optimal level of 1 mM Mg2+ or 2 mM Mn2+, respectively
Mg2+
-
this enzyme is composed of two dissociable subunits that exhibit a catalytic activity only when they are associated together in the presence of a cofactor, Mg2+, and a substrate, farnesyl diphosphate. The quartz-crystal microbalance measurement reveals that farnesyl diphosphate is preferentially bound to subunit II in the presence of Mg2+, while the atomic force microscopy measurement shows that the adhesive force between the subunits is observed only in the presence of both Mg2+ and farnesyl diphosphate
Mn2+
P31112 and P31114
30% of the activation obtained with Mg2+
Mn2+
-
requires a divalent cation for the enzymatic activity with an optimal level of 1 mM Mg2+ or 2 mM Mn2+, respectively
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
enzymatic activity is inhibited by each antiserum against component I or component II, indicating that either component I or component II is confirmed immunochemically to be essential for enzymatic activity
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0053
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme L107S
0.0056
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme E128V
0.0064
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme D73A
0.0069
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme D97A
0.0071
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 30C
0.0072
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme K130I; pH 8.5, 37C, mutant enzyme L102S; pH 8.5, 37C, wild-type enzyme
0.0077
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, wild-type enzyme
0.0082
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme N127A
0.0084
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme T76V
0.0085
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-B(A79F)
0.0098
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme Y103S
0.0103
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme S100G
0.0116
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-B(I76G)
0.0124
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(A79L)
0.0133
-
(2E,6E)-farnesyl diphosphate
-
pH 7.5, 37C
0.0133
-
(2E,6E)-farnesyl diphosphate
P31112 and P31114
pH 7.5, 37C
0.02
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme Y104S
0.0508
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme L94S
0.0741
-
(2E,6E)-farnesyl diphosphate
-
pH 8.5, 37C, mutant enzyme V93G
0.0083
-
geranylgeranyl diphosphate
-
pH 7.5, 37C
0.0083
-
geranylgeranyl diphosphate
P31112 and P31114
pH 7.5, 37C
0.0085
-
geranylgeranyl diphosphate
-
pH 8.5, 30C
0.0168
-
geranylgeranyl diphosphate
-
pH 8.5, 37C, wild-type enzyme
0.0198
-
geranylgeranyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(I76G)
0.0344
-
geranylgeranyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(A79L)
0.0568
-
geranylgeranyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(A79F)
0.0128
-
isopentenyl diphosphate
-
pH 7.5, 37C
0.0128
-
isopentenyl diphosphate
P31112 and P31114
pH 7.5, 37C
0.0134
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme K130I
0.0142
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme S100G
0.0143
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(A79F)
0.0156
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme D97A
0.0161
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme D73A
0.0167
-
isopentenyl diphosphate
-
pH 8.5, 30C
0.0167
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme Y104S; pH 8.5, 37C, wild-type enzyme
0.0172
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme E128V
0.0175
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme L107S
0.0183
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme T76V
0.0188
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme Y103S
0.0194
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme L94S; pH 8.5, 37C, mutant enzyme N127A
0.0198
-
isopentenyl diphosphate
-
pH 8.5, 37C, wild-type enzyme
0.0203
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme L102S
0.0214
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(I76G)
0.0226
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme V93G
0.0259
-
isopentenyl diphosphate
-
pH 8.5, 37C, mutant enzyme subunit-I(wild-type)/subunit-II(A79L)
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
7
P31112 and P31114
-
7.5
9
-
recombinant enzyme
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
8
-
pH 5.5: about 50% of maximal activity, pH 8.0: about 50% of maximal activity
PDB
SCOP
CATH
ORGANISM
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Pseudoalteromonas atlantica (strain T6c / ATCC BAA-1087)
Pseudoalteromonas atlantica (strain T6c / ATCC BAA-1087)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30000
-
P31112 and P31114
molecular weight of enzyme component I and enzyme component II is both about 30000 Da, gel filtration
45000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
P31112 and P31114
molecular weight of enzyme component I and enzyme component II is both about 30000 Da, gel filtration
?
-
x * 24610 (subunit 1) + x * 36172 (subunit 2), calculated from sequence
?
Bacillus subtilis PCI-219
-
molecular weight of enzyme component I and enzyme component II is both about 30000 Da, gel filtration
-
dimer
-
1 * 29000 (enzyme component I) + 1 * 36000 (enzyme component II), SDS-PAGE
dimer
-
this enzyme is composed of two dissociable subunits that exhibit a catalytic activity only when they are associated together in the presence of a cofactor, Mg2+, and a substrate, farnesyl diphosphate. The quartz-crystal microbalance measurement reveals that farnesyl diphosphate is preferentially bound to subunit II in the presence of Mg2+, while the atomic force microscopy measurement shows that the adhesive force between the subunits is observed only in the presence of both Mg2+ and farnesyl diphosphate
heterodimer
P31112 and P31114
1 * 29122 (GerC1) + 1 * 39516 (GerC3), GerC3 supplies the sites for the substrate binding and catalytic activity of HepPP synthase, GerC1 plays an auxiliary but essential role in enzymatic catalysis, calculated from sequence
additional information
-
the hybrid-type combination of component I (Bacillus subtilis) and component II (Bacillus stearothermophilus) gives distinct prenyltransferase activity. The hybrid-type enzyme catalyzes the synthesis of heptaprenyl diphosphate and shows moderate heat stability, which is between those of the natural enzymes from Bacillus subtilis and Bacillus stearothermophilus. There is no possibility of forming a hybrid between the heptaprenyl and hexaprenyl diphosphate synthases
additional information
-
the hybrid-type combination of component I (Bacillus subtilis) and component II (Bacillus stearothermophilus) gives distinct prenyltransferase activity. The hybrid-type enzyme catalyzes the synthesis of heptaprenyl diphosphate and shows moderate heat stability, which is between those of the natural enzymes from Bacillus subtilis and Bacillus stearothermophilus. There is no possibility of forming a hybrid between the heptaprenyl and hexaprenyl diphosphate synthases
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
50
-
P31112 and P31114
component II is inactivated almost completely within 5 min, component II does not lose activity within 30 min
50
-
-
30 min, wild-type and mutant enzymes retain almost original activity after heating
additional information
-
-
the hybrid-type combination of component I (Bacillus subtilis) and component II (Bacillus stearothermophilus) shows moderate heat stability, which is between those of the natural enzymes from Bacillus subtilis and Bacillus stearothermophilus
additional information
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the hybrid-type combination of component I (Bacillus subtilis) and component II (Bacillus stearothermophilus) shows moderate heat stability, which is between those of the natural enzymes from Bacillus subtilis and Bacillus stearothermophilus. There is no possibility of forming a hybrid between the heptaprenyl and hexaprenyl diphosphate synthases
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
separation of component I and component II
P31112 and P31114
the two subunits (subunit I and subunit II) of the HepPP synthase are overproduced in Escherichia coli cells respectively and purified. These subunits are modified with six histidines (polyhistidine) at the N-terminal
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to explore the dynamic interaction of the two dissociable components during catalysis, expression vector systems for the two structural genes, gerC1 and gerC3, are constructed separately, and the two components are overproduced in Escherichia coli cells. Each component is purified homogeneously
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Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
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expression in Escherichia coli, mutant enzymes
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the two subunits (subunit I and subunit II) of the HepPP synthase are overproduced in Escherichia coli cells respectively and purified. These subunits are modified with six histidines (polyhistidine) at the N-terminal
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to explore the dynamic interaction of the two dissociable components during catalysis, expression vector systems for the two structural genes, gerC1 and gerC3, are constructed separately, and the two components are overproduced in Escherichia coli cells
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A79F
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mutant enzyme subunit-I(wild-type)/subunit-B(A79F). Comparable kinetic constants with those of the wild-type enzyme. Significantly different pattern of product distribution from that of the wild type. The major product is hexaprenyl diphosphate
A79L
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mutant enzyme subunit-I(wild-type)/subunit-B(A79L). Comparable kinetic constants with those of the wild-type enzyme. Significantly different pattern of product distribution from that of the wild type. The major product is hexaprenyl diphosphate
A79Y
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single replacement to the aromatic residue at the fourth or the fifth position before the first aspartate-rich motif (FARM), mainly yields a C20 product
D73A
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mutant shows similar Km-value for farnesyl diphosphate and Vmax-value compared to the wild-type
D97A
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reaction products have marked differences in chain length distribution from the wild-type enzyme. D97A produces larger amounts of shorter chain prenyl diphosphates
D97A
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mutant enzyme subunit-I(D97A)/subunit-II(wild-type), major products: geranylgeranyl diphosphate, farnesylgeranyl diphosphate, hexaprenyl diphosphate, heptaprenyl diphosphate
D97A/A79F
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mutant enzyme subunit-I(97A)/subunit-II(A79F) forms exclusively geranylgeranyl diphosphate
D97A/A79L
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mutant enzyme subunit-I(D97A)/subunit-II(A79L), the final product is farnesylgeranyl diphosphate, increase in the production of geranylgeranyl diphosphate
E128V
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mutant shows similar Km-value for farnesyl diphosphate and Vmax-value compared to the wild-type
I76G
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can catalyze condensations of isopentenyl diphosphate beyond the native chain length of C35. With farnesyl diphosphate as allylic substrate the mutant enzyme largely produces (all-trans)-octaprenyl diphosphate (C40, 21%) with a small amount of solanesyl diphosphate (C45, 5.3%). With geranylgeranyl diphosphate as allylic substrate the mutant enzyme yields solanesyl diphosphate (33%) as the main product with a large amount of (all-trans)-octaprenyl diphosphate (21%)
K130I
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mutant shows similar Km-value for farnesyl diphosphate and Vmax-value compared to the wild-type
N127A
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mutant shows similar Km-value for farnesyl diphosphate and Vmax-value compared to the wild-type
S80F
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single replacement to the aromatic residue at the fourth or the fifth position before the first aspartate-rich motif (FARM), mainly yields a C20 product
T76V
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mutant shows similar Km-value for farnesyl diphosphate and Vmax-value compared to the wild-type
V76I
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mutant enzyme subunit-I(wild-type)/subunit-II(A79L), octaprenyl diphosphate is the final product with farnesyl diphosphate as allylic primer
V93G
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16fold lower Vmax values and 10fold higher Km-values for the allylic substrate farnesyl diphosphate compared to wild-type enzyme
Y103S
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reaction products have marked differences in chain length distribution from the wild-type enzyme. Y103S gives octaprenyl diphosphate (C40) as the final product
Y103S
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mutant enzyme subunit-I(Y103S)/subunit-II(wild-type), final product is octaprenyl diphosphate
L94S
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9fold lower Vmax values and 7fold higher Km-values for the allylic substrate farnesyl diphosphate compared to wild-type enzyme
additional information
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the hybrid-type combination of component I (Bacillus subtilis) and component II' (Bacillus stearothermophilus) gives distinct prenyltransferase activity. The hybrid-type enzyme catalyzes the synthesis of heptaprenyl diphosphate and shows moderate heat stability, which is between those of the natural enzymes from Bacillus subtilis and Bacillus stearothermophilus
additional information
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several amino acid residues in the larger subunits Bacillus subtilis heptaprenyl diphosphate synthase are selected for substitutions by site-directed mutagenesis and examined by combination with the corresponding wild type or mutated smaller subunits
additional information
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elucidation of the mechanism of chain length determination of Bacillus stearothermophilus HepPS by site-directed mutagenesis
Y104S
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5fold lower Vmax values and 3fold higher Km-values for the allylic substrate farnesyl diphosphate compared to wild-type enzyme
additional information
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the hybrid-type combination of component I (Bacillus subtilis) and component II (Bacillus stearothermophilus) gives distinct prenyltransferase activity. The hybrid-type enzyme catalyzes the synthesis of heptaprenyl diphosphate and shows moderate heat stability, which is between those of the natural enzymes from Bacillus subtilis and Bacillus stearothermophilus. There is no possibility of forming a hybrid between the heptaprenyl and hexaprenyl diphosphate synthases