Literature summary for 4.2.3.131 extracted from

Brueckner, K.; Bozic, D.; Manzano, D.; Papaefthimiou, D.; Pateraki, I.; Scheler, U.; Ferrer, A.; de Vos, R.C.; Kanellis, A.K.; Tissier, A.
Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes (2014), Phytochemistry, 101, 52-64 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene SKL1, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, RoKSL1 and RoKSL2 cluster into a discrete clade, together with other KSL proteins from the Lamiaceae and Solanaceae (NtABS, ShSBS and SlPHS). Functional recombinant expression in Saccharomyces cerevisiae strain AM113, and transient coexpression of RoSKL1 and RoSKL2 in Nicotiana benthamiana leaves	Salvia rosmarinus
gene SKL2, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, RoKSL1 and RoKSL2 cluster into a discrete clade, together with other KSL proteins from the Lamiaceae and Solanaceae (NtABS, ShSBS and SlPHS). Functional recombinant expression in Saccharomyces cerevisiae strain AM113, and transient coexpression of RoSKL1 and RoSKL2 in Nicotiana benthamiana leaves	Salvia rosmarinus

Cloned (Comment)

Organism

gene SKL1, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, RoKSL1 and RoKSL2 cluster into a discrete clade, together with other KSL proteins from the Lamiaceae and Solanaceae (NtABS, ShSBS and SlPHS). Functional recombinant expression in Saccharomyces cerevisiae strain AM113, and transient coexpression of RoSKL1 and RoSKL2 in Nicotiana benthamiana leaves

Salvia rosmarinus

gene SKL2, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, RoKSL1 and RoKSL2 cluster into a discrete clade, together with other KSL proteins from the Lamiaceae and Solanaceae (NtABS, ShSBS and SlPHS). Functional recombinant expression in Saccharomyces cerevisiae strain AM113, and transient coexpression of RoSKL1 and RoSKL2 in Nicotiana benthamiana leaves

Salvia rosmarinus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
geranylgeranyl diphosphate	Salvia rosmarinus	-	(+)-copalyl diphosphate	-	r
additional information	Salvia rosmarinus	substrate specificity analysis, rosemary RoKSL1/2 are functional enzymes able to convert either 8-OH-CDP or CDP into manoyl oxide or miltiradiene, respectively, and that they likely function in planta as miltiradiene synthases	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Salvia rosmarinus	W8QEG7	-	-
Salvia rosmarinus	W8QMF8	-	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
leaf	young and old	Salvia rosmarinus	-
additional information	quantitative RT-PCR expression analysis on whole leaves shows that RoKSL2 and RoKSL1 are preferentially expressed in young leaves, with RoKSL1 being much less expressed than RoKSL2. Direct comparison of the expression levels between RoKSL1 and RoKSL2 in isolated trichomes and in the remaining leaves indicates that RoKSL1 is not specific to the trichomes and significantly less expressed than RoKSL2 (RoKSL1 transcript level about 1% of expression of RoKSL2)	Salvia rosmarinus	-
trichome	glandular	Salvia rosmarinus	-
trichome	glandular, very limited expression level	Salvia rosmarinus	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(+)-copalyl diphosphate	-	Salvia rosmarinus	miltiradiene + diphosphate	-	?
8-hydroxy-(+)-copalyl diphosphate	-	Salvia rosmarinus	manoyl oxide + diphosphate	-	?
geranylgeranyl diphosphate	-	Salvia rosmarinus	(+)-copalyl diphosphate	-	r
additional information	substrate specificity analysis, rosemary RoKSL1/2 are functional enzymes able to convert either 8-OH-CDP or CDP into manoyl oxide or miltiradiene, respectively, and that they likely function in planta as miltiradiene synthases	Salvia rosmarinus	?	-	?

Synonyms

Synonyms	Comment	Organism
RoSKL1	-	Salvia rosmarinus
RoSKL2	-	Salvia rosmarinus

General Information

General Information	Comment	Organism
metabolism	several stereoisomers of copalyl diphosphate (CDP) are found, among which the most common is ent-CDP, the precursor of gibberellins. Other isomers are syn-CDP and normal-CDP, the latter being utilized for the biosynthesis of diterpene resin acids in Gymnosperms and abietane-type tanshinones in Salvia miltiorrhiza. The second step of the cyclization is initiated through the formation of a carbocation upon ionization of the diphosphate linkage of CDP. This is catalyzed by class I terpene synthases which can transform CDP into a variety of polycyclic diterpenoids. By analogy to other labdane-type diterpenoids, the biosynthesis of the diterpene precursor of the carnosic acid pathway in rosemary proceeds in two sequential steps catalyzed by two distinct enzymes. The first is a class II TPS enzyme, CPS1, EC 5.5.1.12, that yields normal CDP, which then is converted by a KSL enzyme to yield an abietane diterpene, miltiradiene. Miltiradiene synthase appears in two isoforms, RoKSL1 and RoKSL2, in Rosmarinus officinalis. Hypothetical biosynthetic pathway of carnosic acid and carnosol produced in Rosmarinus officinalis, overview	Salvia rosmarinus