gene KSL6, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged codon-optimized KSL6 in Escherichia coli strain Tuner(DE3)

Isodon rubescens

Mg2+

required

Isodon rubescens

(+)-copalyl diphosphate

Isodon rubescens

isopimara-7,15-diene + diphosphate

?

Isodon rubescens

A0A1Z3GCD1

recombinant His-tagged KSL6 from Escherichia coli strain Tuner(DE3) by nickel affinity chromatography

Isodon rubescens

leaf

Isodon rubescens

(+)-copalyl diphosphate

743504

Isodon rubescens

isopimara-7,15-diene + diphosphate

?

GC-MS analysis of compounds

743504

Isodon rubescens

?

?

IrKSL6

Isodon rubescens

kaurene synthase 6

Isodon rubescens

KSL6

Isodon rubescens

30

assay at

Isodon rubescens

8

assay at

Isodon rubescens

gene KSL6, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged codon-optimized KSL6 in Escherichia coli strain Tuner(DE3)

Isodon rubescens

Mg2+

required

Isodon rubescens

(+)-copalyl diphosphate

Isodon rubescens

isopimara-7,15-diene + diphosphate

?

recombinant His-tagged KSL6 from Escherichia coli strain Tuner(DE3) by nickel affinity chromatography

Isodon rubescens

leaf

Isodon rubescens

(+)-copalyl diphosphate

743504

Isodon rubescens

isopimara-7,15-diene + diphosphate

?

GC-MS analysis of compounds

743504

Isodon rubescens

?

?

30

assay at

Isodon rubescens

8

assay at

Isodon rubescens

evolution

IrKSL3 and IrKSL6 have the gammabetaalpha tridomain structure, as these proteins tend to possess the bidomain structure of IrKSL1, highlighting the evolutionary history of KSL gene domain loss and further elucidating chemical diversity evolution from a macroevolutionary stance in Lamiaceae. Identified IrCPS4 is predicted to be involved in oridonin biosynthesis, 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-CPP mediated biosynthesis. The tridomain enzymes (IrKSL3 and IrKSL6) belong to an ancestral KSL clade that has the loss of g domain, a widespread occurrence in the Lamiaceae. Evolutionary origin of the gamma-domain loss

Isodon rubescens

metabolism

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-CP, synthesized by (+)-copalyl diphosphate synthases IrCPS1 or IrCPS2 (EC 5.5.1.12) to produce isopimaradiene. Enzyme IrKSL2 generates a product with identical EI mass spectrum to isopimaradiene (isopimara-7,15-diene), from ent-copalyl diphosphate

Isodon rubescens

evolution

IrKSL3 and IrKSL6 have the gammabetaalpha tridomain structure, as these proteins tend to possess the bidomain structure of IrKSL1, highlighting the evolutionary history of KSL gene domain loss and further elucidating chemical diversity evolution from a macroevolutionary stance in Lamiaceae. Identified IrCPS4 is predicted to be involved in oridonin biosynthesis, 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-CPP mediated biosynthesis. The tridomain enzymes (IrKSL3 and IrKSL6) belong to an ancestral KSL clade that has the loss of g domain, a widespread occurrence in the Lamiaceae. Evolutionary origin of the gamma-domain loss

Isodon rubescens

metabolism

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-CP, synthesized by (+)-copalyl diphosphate synthases IrCPS1 or IrCPS2 (EC 5.5.1.12) to produce isopimaradiene. Enzyme IrKSL2 generates a product with identical EI mass spectrum to isopimaradiene (isopimara-7,15-diene), from ent-copalyl diphosphate

Isodon rubescens

748238

Gnanasekaran

Heterologous expression of th ...

J. Biol. Eng.

663127

Martin

Functional characterization of ...

Plant Physiol.