Information on EC 5.5.1.20 - prosolanapyrone-III cycloisomerase

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

EC NUMBER
COMMENTARY hide
5.5.1.20
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RECOMMENDED NAME
GeneOntology No.
prosolanapyrone-III cycloisomerase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
prosolanapyrone III = (-)-solanapyrone A
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cyclization
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the Diels-Alder reaction is a [4+2] cycloaddition reaction in which a cyclohexene ring is formed between a 1,3-diene and an electron-deficient alkene via a single pericyclic transition state
Diels-Alder reaction
SYSTEMATIC NAME
IUBMB Comments
prosolanapyrone-III:(-)-solanapyrone A isomerase
The enzyme is involved in the biosynthesis of the phytotoxin solanapyrone in some fungi. The bifunctional enzyme catalyses the oxidation of prosolanapyrone II and the subsequent Diels Alder cycloisomerization of the product prosolanapyrone III to (-)-solanapyrone A (cf. EC 1.1.3.42, prosolanapyrone II oxidase).
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene spnF
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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the Diels-Alder reaction is the key transformation in the biosynthesis of many cyclohexene-containing secondary metabolites. Cyclase SpnF catalyses a transannular [4+2] cycloaddition to form the cyclohexene ring in spinosyn A, it catalyzes the cyclization step of macrolactone
physiological function
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the enzyme is involved in biosynthesis of spinosyn A, a tetracyclic polyketide-derived insecticide from Saccharopolyspora spinosa
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(3E,5E,7E,10R,12E,14E,17R,18S,22S)-22-ethyl-10,18-dihydroxy-17-methyloxacyclodocosa-3,5,7,12,14-pentaene-2,16-dione
(2R,3aS,5aS,6E,10S,14S,15R,17E,18aS,18bR)-10-ethyl-2,14-dihydroxy-15-methyl-2,3,3a,5a,10,11,12,13,14,15,18a,18b-dodecahydro-1H-indeno[5,4-e]oxacyclopentadecine-8,16-dione
show the reaction diagram
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via intermediate 6, the SpnF-catalysed endomode syn-addition of an alkenyl to a dienyl functionality seems consistent with a Diels-Alder reaction, but a stepwise [4+2] cycloaddition mechanism, for example, one involving a dipolar intermediate such as 7, cannot be ruled out
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-
?
prosolanapyrone III
(-)-solanapyrone A
show the reaction diagram
additional information
?
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mode of formation of the complex perhydro-as-indacene moiety in spinosyn A, overview. No activity with keto-intermediate 3
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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
prosolanapyrone III
(-)-solanapyrone A
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
flavin
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flavin-dependent oxidase, the enzyme possesses a flavin-binding domain
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
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SpnF specifically accelerates the ring formation reaction with an estimated 500fold rate enhancement. The cyclization event is modelled as the sum of a first order and a Michaelis-Menten process and fits
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.23
(3E,5E,7E,10R,12E,14E,17R,18S,22S)-22-ethyl-10,18-dihydroxy-17-methyloxacyclodocosa-3,5,7,12,14-pentaene-2,16-dione
Saccharopolyspora spinosa
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pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.69
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pH and temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
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pH 5.0: about 55% of maximal activity, pH 9.0: about 55% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.25
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isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46000 - 60000
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gel filtration
53000
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gel filtration
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.7
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irreversible inactivation below
714292
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-84°C, the enzyme retains 69% and 86% of its pre-storage activities for the Diels-Alder reaction and oxidation after 5 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3)
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Aspergillus oryzae under the starch inducible alpha-amylase promoter, expression in Pichia pastoris
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gene spnF, overexpression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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