Information on EC 2.5.1.62 - chlorophyll synthase

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

EC NUMBER
COMMENTARY hide
2.5.1.62
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RECOMMENDED NAME
GeneOntology No.
chlorophyll synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
chlorophyllide a + phytyl diphosphate = chlorophyll a + diphosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
polyprenyl-group transfer
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
chlorophyll a biosynthesis I
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chlorophyll a biosynthesis II
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chlorophyll a biosynthesis III
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chlorophyll cycle
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chlorophyll metabolism
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Porphyrin and chlorophyll metabolism
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Metabolic pathways
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
chlorophyllide-a:phytyl-diphosphate phytyltransferase
Requires Mg2+. The enzyme is modified by binding of the first substrate, phytyl diphosphate, before reaction of the modified enzyme with the second substrate, chlorophyllide a, can occur. The reaction also occurs when phytyl diphosphate is replaced by geranylgeranyl diphosphate.
CAS REGISTRY NUMBER
COMMENTARY hide
9077-08-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
enhancer trap line J1511 with yeast transcription factor GAL4-VP16
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Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
L., line 2-24, albina mutant and wild type line 3629
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-
Manually annotated by BRENDA team
cv. Haisa, barley mutant albostrians
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-
Manually annotated by BRENDA team
cultivar Zhenhui 249
UniProt
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
PCC 6803
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-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
enzyme catalyzes the last step in the chlorophyll biosynthetic pathway; enzyme catalyzes the last step in the chlorophyll biosynthetic pathway
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
chlorophyllide + phytyl diphosphate
chlorophyll + diphosphate
show the reaction diagram
chlorophyllide a + farnesyl diphosphate
3-farnesylchlorophyllide a + diphosphate
show the reaction diagram
chlorophyllide a + geranylgeranyl diphosphate
geranylgeranylchlorophyllide a + diphosphate
show the reaction diagram
chlorophyllide a + phytyl diphosphate
chlorophyll a + diphosphate
show the reaction diagram
chlorophyllide a + tetraprenyl diphosphate
diphosphate + 3-tetraprenylchlorophyllide a
show the reaction diagram
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ping-pong mechanism. Tetraprenyl diphosphate must bind to the enzyme as the first substrate and esterification occurs when the pre-loaded enzyme meets the second substrate
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-
?
phenylamino-Zn-pheophorbide b + phytyl diphosphate
?
show the reaction diagram
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-
-
-
?
pheophorbide a + geranylgeranyl diphosphate
?
show the reaction diagram
Zn chlorophyllide a + geranylgeranyl diphosphate
Zn chlorophyll a + diphosphate
show the reaction diagram
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-
-
?
Zn chlorophyllide a + phytyl diphosphate
Zn chlorophyll a + diphosphate
show the reaction diagram
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-
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?
Zn-chlorophyllide a + phytyl diphosphate
Zn-chlorophyll a + diphosphate
show the reaction diagram
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Zn-chlorophyllide a possesses a central Zn2+ ion instead of Mg2+, can be used as as the substrate for chlorophyll synthetase in briefly illuminated etiolated rye leaves
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-
?
Zn-pheophorbide a + phytyl diphosphate
?
show the reaction diagram
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-
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-
?
Zn-pheophorbide b + phytyl diphosphate
?
show the reaction diagram
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-
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
chlorophyllide a + geranylgeranyl diphosphate
geranylgeranylchlorophyllide a + diphosphate
show the reaction diagram
chlorophyllide a + phytyl diphosphate
chlorophyll a + diphosphate
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cerium
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cerium addition significantly relieves the inhibition of chlorophyll biosynthesis in maize caused by magnesium deficiency and increases chlorophyll content. cerium may partly substitue for magnesium
Magnesium
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magnesium deficiency leads to decrease of chlorophyll contents in maize leaves, with concomitant decrease in the expression levels of magnesium chelatase, magnesium-protoporphyrin IX methyltransferase, and chlorophyll synthase. Magnesium deficiency significantly inhibits the transformation from coproporphyrinogen III or protoporphyrin IX to chlorophyll. Cerium addition significantly relieves the inhibition of chlorophyll biosynthesis in maize caused by magnesium deficiency and increases chlorophyll content
Mn2+
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partially reconstitutes activity after treatment with EDTA
Zn2+
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partially reconstitutes activity after treatment with EDTA
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bacteriochlorophyllide a
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competitive inhibitor, effect is reversed by an increased level of chlroophyllide a, no cell growth of 50 microM
diacetyl
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N-Phenylmaleimide
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since the wild-type enzyme and all other Cys-mutants with the exception of the mutant C304A are inhibited, it is concluded that the inhibitor binds to a non-essential Cys residue to abolish activity
additional information
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hairpin structure in plasmid is tran-activated by the yeast transcription factor that is active in veins of leaves and petiols leads to post-transcriptional silencing of the enzyme by RNA interference
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
geranylgeranyl diphosphate
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lysate mixed into reaction buffer (120 mM potassium acetate, 10 mM magnesium acetate, 50 mM HEPES-KOH, pH 7.6, 14 mM 2-mercaptoethanol, 10% glycerol, and 0.5 mM ATP), 30°C
0.32
phytyl diphosphate
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lysate mixed into reaction buffer (120 mM potassium acetate, 10 mM magnesium acetate, 50 mM HEPES-KOH, pH 7.6, 14 mM 2-mercaptoethanol, 10% glycerol, and 0.5 mM ATP), 30°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.3 - 1.14
bacteriochlorophyllide a
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6
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assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26
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assay at
27
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assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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of wild-type line 3629
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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peripheral membrane protein which releases its product into the membrane
Manually annotated by BRENDA team
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inner membrane fraction
Manually annotated by BRENDA team
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chlorophyll synthetase activity is relocated from transforming prolamellar bodies to developing thylakoids during irradiation of dark-grown Triticum aestivum
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
calculated; calculated
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cell harvesting, washing with 10 mM Tris-HCl, pH 8.0, freeze-thaw-cycles, incubated with DNase and 5 mM MgCl2 at room temperature, lysate used for assays
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
amplified by the polymerase chain reaction and cloned into T7 RNA polymerase-based expression plasmid, heterologous expression of chlG in Escherichia coli
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expression in Escherichia coli
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hairpin structure is cloned into Binary plasmid, plants are transformed via Agrobacterium tumefaciens
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PCR-amplification, in T-vector, expression plasmid pRSET-A in Escherichia coli BL21(DE3)/pLysS
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subcloning in the pGEM-T vector, cloning in the pBirAR vector for overexpression or gene silencing, transformation of Nicotiana tabacum leaf discs mediated by Agrobacterium tumefaciens strain GV2260; subcloning in the pGEM-T vector, cloning in the pBirAR vector for overexpression or gene silencing, transformation of Nicotiana tabacum leaf discs mediated by Agrobacterium tumefaciens strain GV2260
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
magnesium deficiency leads to decrease of chlorophyll contents in maize leaves, with concomitant decrease in the expression levels of magnesium chelatase, magnesium-protoporphyrin IX methyltransferase, and chlorophyll synthase
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C109A
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mutant enzyme exhibits nearly no enzymatic activity, N-phenylmaleimide results in an additional decrease of activity
C130A
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mutant enzyme shows reduced activity and sensitivity to N-phenylmaleimide
C137A
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mutant enzyme is not impaired in enzymatic activity and shows the same inhibition by N-phenylmaleimide as the wild-type enzyme
C262A
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mutant enzyme is not impaired in enzymatic activity and shows the same inhibition by N-phenylmaleimide as the wild-type enzyme
C304A
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as active as the wild-type enzyme, mutant enzyme is not inhibited by N-phenylmaleimide
D147A
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mutant enzyme without activity
D150A
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mutant enzyme without activity
D154A
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mutant enzyme without activity
N146A
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mutant enzyme without activity
R151A
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mutant enzyme with 35% of the activity compared to wild-type enzyme
R161A
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mutant enzyme without activity
R161H
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mutant enzyme with 1% activity compared to wild type enzyme
R161K
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mutant enzyme with 34% activity compared to wild type enzyme
R284A
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mutant enzyme shows wild-type activity
R91A
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mutant enzyme without activity
P198S
the chlorophyll-deficient mutant yellow-green leaf1 (ygl1) with amino acid substitution P198S shows yellow-green leaves in young plants with decreased chlorophyll synthesis, increased level of tetrapyrrole intermediates, and delayed chloroplast development, and exhibits approximately 35.22% and 21.75% esterification of chlorophyllide a with geranylgeranly diphosphate and phytyl diphosphate, respectively, compared to wild type recombinant enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
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