Literature summary for 2.5.1.67 extracted from

Yang, T.; Gao, L.; Hu, H.; Stoopen, G.; Wang, C.; Jongsma, M.A.
Chrysanthemyl diphosphate synthase operates in planta as a bifunctional enzyme with chrysanthemol synthase activity (2014), J. Biol. Chem., 289, 36325-36335.

Cloned(Commentary)

Cloned (Comment)	Organism
gene CDS, dNA and amino acid sequence determination and analysis, sequence comparisons, recombinant CDS overexpression under the control of chrysanthemum RbcS1 promoter in transgenic Nicotiana tabacum cv. Samsun plants, in leaf chloroplasts, using Agrobacterium tumefaciens strain AGL-0 transfection method, the expression leads to emittation of chrysanthemol. Recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(AI)	Tanacetum cinerariifolium

Cloned (Comment)

Organism

gene CDS, dNA and amino acid sequence determination and analysis, sequence comparisons, recombinant CDS overexpression under the control of chrysanthemum RbcS1 promoter in transgenic Nicotiana tabacum cv. Samsun plants, in leaf chloroplasts, using Agrobacterium tumefaciens strain AGL-0 transfection method, the expression leads to emittation of chrysanthemol. Recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(AI)

Tanacetum cinerariifolium

Protein Variants

Protein Variants	Comment	Organism
N283D	site-directed mutagenesis, replacing the asparagine with either aspartate or glycine in the aspartate-rich motif abolishes the enzyme's terpene synthase and prenyltransferase activities	Tanacetum cinerariifolium
N283G	site-directed mutagenesis, replacing the asparagine with either aspartate or glycine in the aspartate-rich motif abolishes the enzyme's terpene synthase and prenyltransferase activities	Tanacetum cinerariifolium

Inhibitors

Inhibitors	Comment	Organism	Structure
dimethylallyl diphosphate	DMAPP, causes significant substrate inhibition at elevated concentrations for chrysanthemol synthesis, increase of DMAPP concentration leads to a decrease of chrysanthemol but a strong further increase of chrysanthemyl diphosphate production	Tanacetum cinerariifolium

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Tanacetum cinerariifolium

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
chloroplast	the N-terminal peptide of enzyme CDS is a plastid-targeting peptide	Tanacetum cinerariifolium	9507	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Tanacetum cinerariifolium

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2 dimethylallyl diphosphate	Tanacetum cinerariifolium	-	diphosphate + chrysanthemyl diphosphate	-	?
additional information	Tanacetum cinerariifolium	chrysanthemyl diphosphate synthase operates in planta as a bifunctional enzyme with chrysanthemol synthase activity. The NDXXD catalytic motif of CDS, under the lower substrate conditions prevalent in plants, also catalyzes the next step, converting CPP into chrysanthemol by hydrolyzing the diphosphate moiety. For the chrysanthemol synthase activity, dimethylallyl diphosphate competes with chrysanthemyl diphosphate as substrate	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Tanacetum cinerariifolium	L7RFF8	i.e. Chrysantemum cinerariifolium	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(AI) by nickel affinity chromatography and ultrafiltration	Tanacetum cinerariifolium

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2 dimethylallyl diphosphate	-	Tanacetum cinerariifolium	diphosphate + chrysanthemyl diphosphate	-	?
2 dimethylallyl diphosphate	the enzyme catalyzes c1'-2-3 cyclopropanation reactions of two molecules of dimethylallyl diphosphate to yield chrysanthemyl diphosphate	Tanacetum cinerariifolium	diphosphate + chrysanthemyl diphosphate	-	?
additional information	chrysanthemyl diphosphate synthase operates in planta as a bifunctional enzyme with chrysanthemol synthase activity. The NDXXD catalytic motif of CDS, under the lower substrate conditions prevalent in plants, also catalyzes the next step, converting CPP into chrysanthemol by hydrolyzing the diphosphate moiety. For the chrysanthemol synthase activity, dimethylallyl diphosphate competes with chrysanthemyl diphosphate as substrate	Tanacetum cinerariifolium	?	-	?
additional information	The enzyme shows activity to produce diphosphates (chrysanthemyl diphosphate and lavandulyl diphosphate) and terpene alcohols (chrysanthemol and lavandulol) from dimethylallyl diphosphate	Tanacetum cinerariifolium	?	-	?

Synonyms

Synonyms	Comment	Organism
CDS	-	Tanacetum cinerariifolium
chrysanthemol synthase	UniProt	Tanacetum cinerariifolium

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Tanacetum cinerariifolium

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Tanacetum cinerariifolium

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the very large family of short-chain prenyltransferases and terpene synthases	Tanacetum cinerariifolium
metabolism	chrysanthemyl diphosphate synthase is the first pathway-specific enzyme in the biosynthesis of pyrethrins	Tanacetum cinerariifolium
additional information	CDS should be renamed a chrysanthemol synthase utilizing dimethylallyl diphosphate as substrate	Tanacetum cinerariifolium
physiological function	chrysanthemyl diphosphate synthase is the first pathway-specific enzyme in the biosynthesis of pyrethrins, the most widely used plant-derived pesticide. The enzyme shows activity to produce diphosphates (chrysanthemyl diphosphate and lavandulyl diphosphate) and terpene alcohols (chrysanthemol and lavandulol) from dimethylallyl diphosphate	Tanacetum cinerariifolium