three KSL genes are involved in ent-CPP interactions, and a normal-CPP mediated miltiradiene and isopimaradiene biosynthesis pathway. The three KSL genes bearing different domain structures are involved in the normal-copalyl diphosphate mediated biosynthesis. IrKSL2, IrKSL4, and IrKSL5 react with ent-copalyl diphosphate (ent-CPP) to produce an ent-isopimaradiene-like compound, ent-atiserene and ent-kaurene, respectively. IrKSL3 and IrKSL6 form a monophyletic clade with IrKSL1 despite the fact that they have the common gammabetaalpha tridomain structures. IrKSL2, IrKSL4, and IrKSL5 belong to the KSLs that are involved in the ent-CPP-mediated biosynthetic pathway that catalyzes reactions with ent-CPP and ent-LDPP
ent-kaurene diterpenoids are the largest group of known Isodon diterpenoids, five copalyl diphosphate synthase (CPS) and six kaurene synthase-like (KSL) genes are identified by transcriptome profiling of Isodon rubescens leaves. IrKSL6 reacts with normal-CPP to produce isopimaradiene. Enzyme IrKSL2 generates a product with identical EI mass spectrum to isopimaradiene (isopimara-7,15-diene), from ent-copalyl diphosphate synthesized by enzyme with IrCPS4, an ent-copalyl diphosphate synthase (EC 126.96.36.199)
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gene KSL2, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged codon-optimized KSL2 in Escherichia coli strain Tuner(DE3)