Literature summary for 1.14.14.176 extracted from

Rontein, D.; Onillon, S.; Herbette, G.; Lesot, A.; Werck-Reichhart, D.; Sallaud, C.; Tissier, A.
CYP725A4 from yew catalyzes complex structural rearrangement of taxa-4(5),11(12)-diene into the yclic ether 5(12)-oxa-3(11)-cyclotaxane (2008), J. Biol. Chem., 283, 6067-6075.

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Application

Application	Comment	Organism
drug development	paclitaxel is a successful anticancer drug	Taxus cuspidata

Cloned(Commentary)

Cloned (Comment)	Organism
cDNAs encoding taxa-4(5),11(12)-diene synthase and CYP725A4 are introduced in Nicotiana sylvestris for specific expression in trichome cells. Production of truncated TS in Escherichia coli. The TS cDNA coding for a protein truncated at the N-terminus at amino acid position 75 is cloned in the pET-30b expression vector. Furthermore, CYP725A4 is expressed in yeast.	Taxus cuspidata
CYP725A4, DNA and amino acid sequence determination and analysis, expression in the Saccharomyces cerevisiae WAT11 strain containing the Arabidopsis reductase ATR1, and co-expression in Nicotiana sylvestris with taxa-4(5),11(12)-diene synthase, which does not lead to 5alpha-hydroxytaxa-4(20),11(12)-diene, but to formation of 5(12)-oxa-3(11)-cyclotaxane, a tritiated oxidized taxane, structural analysis of the compound, phenotype, overview	Taxus cuspidata

Cloned (Comment)

Organism

cDNAs encoding taxa-4(5),11(12)-diene synthase and CYP725A4 are introduced in Nicotiana sylvestris for specific expression in trichome cells. Production of truncated TS in Escherichia coli. The TS cDNA coding for a protein truncated at the N-terminus at amino acid position 75 is cloned in the pET-30b expression vector. Furthermore, CYP725A4 is expressed in yeast.

Taxus cuspidata

CYP725A4, DNA and amino acid sequence determination and analysis, expression in the Saccharomyces cerevisiae WAT11 strain containing the Arabidopsis reductase ATR1, and co-expression in Nicotiana sylvestris with taxa-4(5),11(12)-diene synthase, which does not lead to 5alpha-hydroxytaxa-4(20),11(12)-diene, but to formation of 5(12)-oxa-3(11)-cyclotaxane, a tritiated oxidized taxane, structural analysis of the compound, phenotype, overview

Taxus cuspidata

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0069	-	taxa-4(5),11(12)-diene	-	Taxus cuspidata

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
additional information	Taxus cuspidata	cDNAs encoding taxa-4(5),11(12)-diene synthase and CYP725A4 are introduced in Nicotiana sylvestris for specific expression in trichome cells. The co-expression does not lead to the production of the expected 5-hydroxytaxa-4(20),11(12)-diene. Instead, taxa-4(5),11(12)-diene is quantitatively converted to a novel taxane	?	-	?
taxa-4(5),11(12)-diene + [reduced NADPH-hemoprotein reductase] + O2	Taxus cuspidata	hydroxylation of the C-5alpha is the first of the oxidations resulting in taxol	5(12)-oxa-3(11)-cyclotaxane + [oxidized NADPH-hemoprotein reductase] + H2O	OCT is a cyclic ether	?
taxa-4,11-diene + [reduced NADPH-hemoprotein reductase] + O2	Taxus cuspidata	-	taxa-4(20),11-dien-5alpha-ol + [oxidized NADPH-hemoprotein reductase] + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Taxus cuspidata	Q6WG30	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
taxa-4,11-diene + [reduced NADPH-hemoprotein reductase] + O2 = taxa-4(20),11-dien-5alpha-ol + [oxidized NADPH-hemoprotein reductase] + H2O	reaction mechanism	Taxus cuspidata	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
leaf	-	Taxus cuspidata	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	the recombinant enzyme produces 5(12)-oxa-3(11)-cyclotaxane, a tritiated oxidized taxane, instead of taxa-4(20),11-dien-5alpha-ol, structural analysis of the compound, overview	Taxus cuspidata	?	-	?
additional information	cDNAs encoding taxa-4(5),11(12)-diene synthase and CYP725A4 are introduced in Nicotiana sylvestris for specific expression in trichome cells. The co-expression does not lead to the production of the expected 5-hydroxytaxa-4(20),11(12)-diene. Instead, taxa-4(5),11(12)-diene is quantitatively converted to a novel taxane	Taxus cuspidata	?	-	?
taxa-4(5),11(12)-diene + [reduced NADPH-hemoprotein reductase] + O2	hydroxylation of the C-5alpha is the first of the oxidations resulting in taxol	Taxus cuspidata	5(12)-oxa-3(11)-cyclotaxane + [oxidized NADPH-hemoprotein reductase] + H2O	OCT is a cyclic ether	?
taxa-4,11-diene + [reduced NADPH-hemoprotein reductase] + O2	-	Taxus cuspidata	taxa-4(20),11-dien-5alpha-ol + [oxidized NADPH-hemoprotein reductase] + H2O	-	?

Synonyms

Synonyms	Comment	Organism
CYP725A4	-	Taxus cuspidata
taxa-4(5),11(12)-diene 5-hydroxylase	-	Taxus cuspidata
taxa-4(5),11(12)-diene 5alpha-hydroxylase	-	Taxus cuspidata

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Taxus cuspidata

Cofactor

Cofactor	Comment	Organism	Structure
NADPH	-	Taxus cuspidata