Information on EC 1.3.99.31 - phytoene desaturase (lycopene-forming)

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.3.99.31
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RECOMMENDED NAME
GeneOntology No.
phytoene desaturase (lycopene-forming)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
15-cis-phytoene + 4 acceptor = all-trans-lycopene + 4 reduced acceptor
show the reaction diagram
overall reaction
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15-cis-phytoene + acceptor = all-trans-phytofluene + reduced acceptor
show the reaction diagram
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all-trans-neurosporene + acceptor = all-trans-lycopene + reduced acceptor
show the reaction diagram
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all-trans-phytofluene + acceptor = all-trans-zeta-carotene + reduced acceptor
show the reaction diagram
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all-trans-zeta-carotene + acceptor = all-trans-neurosporene + reduced acceptor
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
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Carotenoid biosynthesis
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
15-cis-phytoene:acceptor oxidoreductase (lycopene-forming)
Requires FAD. The enzyme is involved in carotenoid biosynthesis and catalyses up to four desaturation steps (cf. EC 1.3.99.28 [phytoene desaturase (neurosporene-forming)], EC 1.3.99.29 [phytoene desaturase (zeta-carotene-forming)] and EC 1.3.99.30 [phytoene desaturase (3,4-didehydrolycopene-forming)]).
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene crtI
UniProt
Manually annotated by BRENDA team
gene crtI
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Manually annotated by BRENDA team
gene crtI
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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CRTI-type phytoene desaturases prevailing in bacteria and fungi can form lycopene directly from phytoene while plants employ two distinct desaturases and two cis-tans isomerases for the same purpose
malfunction
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a mutant of Rubrivivax gelatinosus lacking the crtI gene produces only phytoene, indicating that this organism has no other phytoene desaturases. When the crtI deletion mutant is complemented by the three-step phytoene desaturase of Rhodobacter capsulatus, spirilloxanthin and its precursors are not synthesized, although spheroidene and OH-spheroidene are accumulated
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
15-cis-phytoene + 3 acceptor
all-trans-neurosporene + 3 reduced acceptor
show the reaction diagram
15-cis-phytoene + 4 acceptor
all-trans-lycopene + 4 reduced acceptor
show the reaction diagram
15-cis-phytoene + 4 acceptor
all-trans-lycopene + reduced 4 acceptor
show the reaction diagram
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-
-
-
?
15-cis-phytoene + acceptor
all-trans-phytofluene + reduced acceptor
show the reaction diagram
15-cis-phytoene + FAD
all-trans-phytofluene + FADH2
show the reaction diagram
-
-
-
-
?
all-trans-neurosporene + acceptor
all-trans-lycopene + reduced acceptor
show the reaction diagram
all-trans-neurosporene + FAD
all-trans-lycopene + FADH2
show the reaction diagram
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-
-
-
?
all-trans-phytofluene + 3 acceptor
all-trans-lycopene + 3 reduced acceptor
show the reaction diagram
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-
-
-
?
all-trans-phytofluene + acceptor
all-trans-zeta-carotene + reduced acceptor
show the reaction diagram
all-trans-phytofluene + FAD
all-trans-zeta-carotene + FADH2
show the reaction diagram
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-
-
-
?
all-trans-zeta-carotene + 2 acceptor
all-trans-lycopene + 2 reduced acceptor
show the reaction diagram
all-trans-zeta-carotene + acceptor
all-trans-neurosporene + reduced acceptor
show the reaction diagram
all-trans-zeta-carotene + FAD
all-trans-neurosporene + FADH2
show the reaction diagram
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-
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-
?
additional information
?
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at higher concentrations, phytoene is the preferred substrate for CrtI, and neurosporene is produced as the major desaturation product, EC 1.3.99.28. At lower phytoene concentrations, neurosporene can be further desaturated by CrtI to produce lycopene
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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
15-cis-phytoene + 3 acceptor
all-trans-neurosporene + 3 reduced acceptor
show the reaction diagram
15-cis-phytoene + 4 acceptor
all-trans-lycopene + 4 reduced acceptor
show the reaction diagram
15-cis-phytoene + 4 acceptor
all-trans-lycopene + reduced 4 acceptor
show the reaction diagram
-
-
-
-
?
15-cis-phytoene + acceptor
all-trans-phytofluene + reduced acceptor
show the reaction diagram
15-cis-phytoene + FAD
all-trans-phytofluene + FADH2
show the reaction diagram
-
-
-
-
?
all-trans-neurosporene + acceptor
all-trans-lycopene + reduced acceptor
show the reaction diagram
all-trans-neurosporene + FAD
all-trans-lycopene + FADH2
show the reaction diagram
-
-
-
-
?
all-trans-phytofluene + 3 acceptor
all-trans-lycopene + 3 reduced acceptor
show the reaction diagram
-
-
-
-
?
all-trans-phytofluene + acceptor
all-trans-zeta-carotene + reduced acceptor
show the reaction diagram
all-trans-phytofluene + FAD
all-trans-zeta-carotene + FADH2
show the reaction diagram
-
-
-
-
?
all-trans-zeta-carotene + 2 acceptor
all-trans-lycopene + 2 reduced acceptor
show the reaction diagram
all-trans-zeta-carotene + acceptor
all-trans-neurosporene + reduced acceptor
show the reaction diagram
all-trans-zeta-carotene + FAD
all-trans-neurosporene + FADH2
show the reaction diagram
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?
additional information
?
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G3FHJ1
at higher concentrations, phytoene is the preferred substrate for CrtI, and neurosporene is produced as the major desaturation product, EC 1.3.99.28. At lower phytoene concentrations, neurosporene can be further desaturated by CrtI to produce lycopene
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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no activity with FMN, NAD+, and NADP+
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0176 - 17.1
15-cis-phytoene
33.1 - 65.8
all-trans-neurosporene
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
56200
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x * 56200, SDS-PAGE
57000
x * 57000, recombinant CrtI, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His6-tagged enzyme from Escherichia coli by nickel affinity cromatography and gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Escherichia coli. Synthesis of the enzyme by different vectors with high and lower copy numbers or strong and weak promoters results in variable ratios of lycopene and neurosporene formation. The highest amount of enzyme is produced with pUC vectors with high copy number and a strong promoter. This transformant synthesizes the highest lycopene amount. The lowest enzyme concentrations is found with pPEU carrying a weak promoter. In this transformant lycopene formation is minor. With increased enzyme concentration, a proportion of lycopene higher than in Rubrivivax gelatinosus cells can be obtained
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gene crtI, cloning of cluster crtCDEF
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gene crtI, cloning of the crt gene cluster crtCDEF, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene crtI, recombinant coexpression of genes crtE, crtB, and crtI from Erwinia uredovora and of crtL from Ficus carica in Pichia pastoris X-33 strains for large-scale biosynthesis of carotenoids, subcloning of crtI in Escherichhia coli strain TOP10
gene crtI, recombinant expression of His6-tagged enzyme in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L153P/L278P
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increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 12:88
L208F
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increased formation of lycopene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 26:74 after complementation of CrtI in pPEU(DH5a/pACCrtEBEU/pPEUCrtIRg), the ratio of the mutant enzyme is 89:11
L208P
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increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 3:97
T256M/D355GL424P
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increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 0:100
Y44C/D53G/P134L/V395A
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increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 3:97
additional information
Show AA Sequence (115 entries)
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