Literature summary for 1.14.14.58 extracted from

Lee, S.; Badieyan, S.; Bevan, D.R.; Herde, M.; Gatz, C.; Tholl, D.
Herbivore-induced and floral homoterpene volatiles are biosynthesized by a single P450 enzyme (CYP82G1) in Arabidopsis (2010), Proc. Natl. Acad. Sci. USA, 107, 21205-21210.

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Cloned(Commentary)

Cloned (Comment)	Organism
cloned into the yeast expression vector YEp352 under the control of the constitutive alcohol dehydrogenase 1 promoter, expression in yeast	Arabidopsis thaliana
expression in yeast WAT11 cells	Arabidopsis thaliana

Crystallization (Commentary)

Crystallization (Comment)	Organism
molecular docking of (E,E)-geranyllinalool and (E)-nerolidol shows that both substrates occupiy the same position in the enzyme binding site with the hydroxyl group at C-3, forming a strong hydrogen bond to the carbonyl oxygen of Thr313. The position of the allylic hydrogen atoms at C-5 of (E,E)-geranyllinalool and (E)-nerolidol and the hydroxyl group at C-3 relative to the reactive iron-oxo heme moiety supports an oxidative-bond cleavage reaction proceeding by a syn-elimination mechanism	Arabidopsis thaliana

Crystallization (Comment)

Organism

molecular docking of (E,E)-geranyllinalool and (E)-nerolidol shows that both substrates occupiy the same position in the enzyme binding site with the hydroxyl group at C-3, forming a strong hydrogen bond to the carbonyl oxygen of Thr313. The position of the allylic hydrogen atoms at C-5 of (E,E)-geranyllinalool and (E)-nerolidol and the hydroxyl group at C-3 relative to the reactive iron-oxo heme moiety supports an oxidative-bond cleavage reaction proceeding by a syn-elimination mechanism

Arabidopsis thaliana

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.00184	-	(3S,6E)-nerolidol	pH and temperature not specified in the publication	Arabidopsis thaliana
0.00184	-	(3S)-(E)-nerolidol	pH not specified in the publication, temperature not specified in the publication	Arabidopsis thaliana
0.00268	-	(E,E)-geranyllinalool	pH and temperature not specified in the publication	Arabidopsis thaliana
0.00268	-	(3S)-(E,E)-geranyllinalool	pH not specified in the publication, temperature not specified in the publication	Arabidopsis thaliana

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
microsome	-	Arabidopsis thaliana	-	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
(3S)-(E,E)-geranyllinalool + [reduced NADPH-hemoprotein reductase] + O2	Arabidopsis thaliana	-	(E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?
(3S)-(E,E)-geranyllinalool + [reduced NADPH-hemoprotein reductase] + O2	Arabidopsis thaliana	biosynthesis of the volatile organic compound (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene	(E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?
(3S,6E)-nerolidol + [reduced NADPH-hemoprotein reductase] + O2	Arabidopsis thaliana	in vitro activity is 5.3fold higher than with (E,E)-geranyllinalool. (3E)-4,8-dimethylnona-1,3,7-triene is not emitted from Arabidopsis leaves (or only in negligible amounts according to our analysis) because of the absence of a prominent (E)-nerolidol synthase activity in this tissue	(3E)-4,8-dimethylnona-1,3,7-triene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	Q9LSF8	-	-
Arabidopsis thaliana	Q9LSF8	ecotype Columbia	-

Reaction

Reaction	Comment	Organism	Reaction ID
(6E,10E)-geranyllinalool + [reduced NADPH-hemoprotein reductase] + O2 = (3E,7E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene + [oxidized NADPH-hemoprotein reductase] + but-3-en-2-one + 2 H2O	the exact mechanism of enzyme is not yet determined. It is possible that CYP82G1 may promote the direct transformation of (E,E)-geranyllinalool and (3R,6E)-nerolidol to (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene and (3E)-4,8-dimethylnona-1,3,7-triene, respectively, with the concomitant release of but-1-en-3-one. But it is also possible that a two-step conversion occurs that includes the formation of intermediate compounds C18 (E,E)-farnesylacetone (from (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene) and C13 (E)-geranylacetone (from (3E)-4,8-dimethylnona-1,3,7-triene). A C4-cleavage product (but-1-en-3-one) resulting from the breakdown of (E,E)-geranyllinalool or (E)-nerolidol is not observed, neither in vitro nor in vivo, and none of the previously proposed ketone intermediates, C18-farnesylacetone and C13-geranylacetone, are detected	Arabidopsis thaliana	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
inflorescence	constitutive expression	Arabidopsis thaliana	-
inflorescence	constitutively expressed	Arabidopsis thaliana	-
leaf	highly coordinated herbivore-induced expression with geranyllinalool synthase in leaves depending on the F-box protein COI-1	Arabidopsis thaliana	-
stem	constitutive expression	Arabidopsis thaliana	-
stem	constitutively expressed	Arabidopsis thaliana	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(3S)-(E,E)-geranyllinalool + [reduced NADPH-hemoprotein reductase] + O2	-	Arabidopsis thaliana	(E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?
(3S)-(E,E)-geranyllinalool + [reduced NADPH-hemoprotein reductase] + O2	biosynthesis of the volatile organic compound (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene	Arabidopsis thaliana	(E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?
(3S,6E)-nerolidol + [reduced NADPH-hemoprotein reductase] + O2	-	Arabidopsis thaliana	(3E)-4,8-dimethylnona-1,3,7-triene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?
(3S,6E)-nerolidol + [reduced NADPH-hemoprotein reductase] + O2	in vitro activity is 5.3fold higher than with (E,E)-geranyllinalool. (3E)-4,8-dimethylnona-1,3,7-triene is not emitted from Arabidopsis leaves (or only in negligible amounts according to our analysis) because of the absence of a prominent (E)-nerolidol synthase activity in this tissue	Arabidopsis thaliana	(3E)-4,8-dimethylnona-1,3,7-triene + but-1-en-3-one + [oxidized NADPH-hemoprotein reductase] + 2 H2O	-	?
additional information	no activity with isophytol, (E)-geraniol, linalool, 3,7,11-trimethyl-3-dodecanol, (Z)-nerolidol. Less than 1% of the activity with (3S,6E)-nerolidol: (E,E,E)-geranylgeraniol, (E,E)-farnesylacetone, (E,E)-farnesol, (E)-geranylacetone. Construction of a protein model based on multiple mammalian P450 templates with closest sequence similarity. Subsequent molecular docking confirms a position of (E)-nerolidol and (E,E)-geranyllinalool in the active site cavity that allows the suggested oxidative bond cleavage reaction to proceed by a syn-elimination (beta-elimination) mechanism. In particular, the carbonyl oxygen of Thr313 in one of the substrate recognition sites appears to be essential for anchoring the substrates by forming a strong hydrogen bond with the hydroxyl group at C3. The model supported the observed differences in substrate specificities	Arabidopsis thaliana	?	-	?
additional information	cytochrome P450 monooxygenase with narrow substrate specificity for (E,E)-geranyllinalool and (E)-nerolidol. Mechanism follows oxidative bond cleavage of the alcohol substrate via syn-elimination of the polar head, together with an allylic C-5 hydrogen atom. No substrate: racemic linalool or (R)-(-)-linalool	Arabidopsis thaliana	?	-	?

Synonyms

Synonyms	Comment	Organism
At3g25180	-	Arabidopsis thaliana
CYP82G1	-	Arabidopsis thaliana
cytochrome P450 82G1	-	Arabidopsis thaliana
DMNT homoterpene synthase	-	Arabidopsis thaliana
DMNT synthase	-	Arabidopsis thaliana
DMNT/TMTT homoterpene synthase	-	Arabidopsis thaliana
TMTT homoterpene synthase	-	Arabidopsis thaliana
TMTT synthase	-	Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.11	-	(E,E)-geranyllinalool	pH and temperature not specified in the publication	Arabidopsis thaliana
0.11	-	(3S)-(E,E)-geranyllinalool	pH not specified in the publication, temperature not specified in the publication	Arabidopsis thaliana
0.2	-	(3S,6E)-nerolidol	pH and temperature not specified in the publication	Arabidopsis thaliana
0.2	-	(3S)-(E)-nerolidol	pH not specified in the publication, temperature not specified in the publication	Arabidopsis thaliana

Cofactor

Cofactor	Comment	Organism	Structure
cytochrome P450	a cytochrome P450 monooxygenase	Arabidopsis thaliana

Expression

Organism	Comment	Expression
Arabidopsis thaliana	constitutively expressed in Arabidopsis stems and inflorescences	additional information
Arabidopsis thaliana	highly coordinated herbivore-induced expression with geranyllinalool synthase in leaves depending on the F-box protein COI-1. Induction of CYP82G1 in response to feeding by Pseudomonas xylostella larvae and upon infection with Pseudomonas syringae DC3000	up

General Information

General Information	Comment	Organism
malfunction	CYP82G1 gene knockout plants do not produce (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene and their phenotype is complemented by the constitutive expression of CYP82G1	Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
41	-	(E,E)-geranyllinalool	pH and temperature not specified in the publication	Arabidopsis thaliana
109	-	(3S,6E)-nerolidol	pH and temperature not specified in the publication	Arabidopsis thaliana