The enzyme has been shown to occur in castor bean (Ricinus communis) and rice (Oryza sativa). ent-Beyerene is also a product of EC 4.2.3.19 (ent-kaurene synthase) of the moss Physcomitrella patens.
The enzyme appears in viruses and cellular organisms
reaction mechanism via carbocation intermediate, in comparison to ant-kaurene synthesis from ent-copalyl diphosphate (EC 4.2.3.19), mass spectrometric analysis
The enzyme has been shown to occur in castor bean (Ricinus communis) and rice (Oryza sativa). ent-Beyerene is also a product of EC 4.2.3.19 (ent-kaurene synthase) of the moss Physcomitrella patens.
Substrates: recombinant OsKSL2 enzyme catalyzes the cyclization of ent-copalyl diphosphate into ent-beyerene as a major and ent-kaurene as a minor product, cf. EC 4.2.3.19, GC-MS product analysis Products: -
Substrates: 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 Products: -
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 RcKS(L)1 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. RcKS(L)1 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
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
construction of a recombinant gene where GFP is fused to the putative chloroplast transit peptide (cTP) sequence of OsKSL2. The fusion gene is transiently expressed in rice leaf protoplasts. GFP signals from the transformed cells overlap with autofluorescence from chloroplasts, indicating that OsKSL2 is localized to the plastid. The subcellular localization of the recombinant GFP-tagged enzyme with GFP fused to a shorter transit peptide sequence from gene Os04g0612000 (UniProt ID Q0JA81) in protoplast cells does not localize to the plastids, the shorter sequence fails to function as a transit peptide
gene OsKSL2, full-length enzyme DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of GST-tagged enzyme in Escherichia coli strain BL21
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
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