Cloned (Comment) | Organism |
---|---|
isozyme determination and phylogenetic analysis and tree, sequence comparisons, overview. Synthetic RcKS(L) isozymes are truncated to remove the N-terminal plastid-directing transit peptide sequences, they are individually subcloned into compatible expression vectors and each coexpressed with either the geranylgeranyl phosphate synthase (GGPS), or the GGPS along with a CPS (EC 5.5.1.13) for recombinant expression in Escherichia coli | Ricinus communis |
Protein Variants | Comment | Organism |
---|---|---|
A676T | site-directed mutagenesis of isozyme RcKSL2, mutation of the catalytically important residue results in 60fold increased product amount composed of essentially the same mix of ent-trachylobane and ent-kaurene as observed for the wild-type enzyme | Ricinus communis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Ricinus communis | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | isozyme RcKSL2 contains a significant deviation from the usually well-conserved (N/D)Dxx(S/T)xxxE sequence that serves as a secondary Mg2+ binding motif | Ricinus communis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ent-copalyl diphosphate | Ricinus communis | - |
ent-kaur-ene + diphosphate | - |
? | |
ent-copalyl diphosphate | Ricinus communis | - |
ent-trachylobane + diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ricinus communis | B9SIL7 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ent-copalyl diphosphate = ent-kaurene + diphosphate | cyclization mechanism derived from quantum chemical calculation for the formation of ent-kaurene and related diterpenes, overview | Ricinus communis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ent-copalyl diphosphate | - |
Ricinus communis | ent-kaurene + diphosphate | - |
? | |
ent-copalyl diphosphate | - |
Ricinus communis | ent-kaur-ene + diphosphate | - |
? | |
ent-copalyl diphosphate | - |
Ricinus communis | ent-trachylobane + diphosphate | - |
? | |
additional information | the enzyme forms ent-kaurene via 4 consecutive intermediates (i.e. ent-pimarenyl, ent-beyeranyl, ent-trachylobanyl, and ent-kauranyl), three of which can be the precursor for different diterpenes other than ent-kaurene, overview. Enzyme product analysis by GC-MS. No production of ent-beyerene or ent-atiserene by isozymes RcKSL1 and RcKSL2 | Ricinus communis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ent-kaurene synthase B | - |
Ricinus communis |
KS(-like) enzyme | - |
Ricinus communis |
KS(L) | - |
Ricinus communis |
RcKS(L) | - |
Ricinus communis |
RcKS1 | - |
Ricinus communis |
RcKSL1 | - |
Ricinus communis |
RcKSL2 | - |
Ricinus communis |
RCOM_0823080 | - |
Ricinus communis |
General Information | Comment | Organism |
---|---|---|
evolution | plants have extensively diversified their arsenal of labdane-related diterpenoids (LRDs), in part via gene duplication and neo-functionalization of the ancestral ent-kaurene synthase (KS) required for gibberellin metabolism. Ricinus communis contains 4 RcKSL isozymes, molecular phylogenetic analysis indicates that RcKSL1 is significantly more closely related to dicotyl ent-kaurene synthases (KSs), while the other three, RcKSL2-4, cluster separately. RcKSL2-4 are in close proximity to each other, within a region of 65 kb, with RcKSL2 and 4 occurring as a tandem gene pair. RcKSL1 is the only isozyme to react with ent-CPP producing small amounts of ent-kaurene, and is referred to as RcKS1. Both RcKSL2 and RcKSL3 also are selectively reacting with ent-CPP with RcKSL2 producing primarily ent-trachylobane (70%) as well as smaller amounts of ent-kaurene (30%), and RcKSL3 producing ent-sandaracopimaradiene (94%) (EC 4.2.3.29) along with small amounts of ent-labdatriene (3%) and ent-pimaradiene (3%). RcKSL4 seems to be inactive, with no products evident from any substrate, but upon synthesis of a codon-optimized gene for the corrected aa sequence and functional analysis, RcKSL4 selectively reacts with ent-CPP and produces largely ent-beyerene (95%) along with very small amounts of ent-atiserene (4%) and ent-kaurene (1%). The enzymes belong to the terpene synthase family | Ricinus communis |
physiological function | plants have extensively diversified their arsenal of labdane-related diterpenoids (LRDs), in part via gene duplication and neo-functionalization of the ancestral ent-kaurene synthase (KS) required for gibberellin metabolism | Ricinus communis |