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Literature summary extracted from

  • Jin, B.; Cui, G.; Guo, J.; Tang, J.; Duan, L.; Lin, H.; Shen, Y.; Chen, T.; Zhang, H.; Huang, L.
    Functional diversification of kaurene synthase-like genes in Isodon rubescens (2017), Plant Physiol., 174, 943-955 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.3.44 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
4.2.3.B70 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) Isodon rubescens
4.2.3.131 cDNA library screening and miltiradiene DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis Isodon rubescens
4.2.3.185 expression in Escherichia coli Isodon rubescens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.3.44 Mg2+ required Isodon rubescens
4.2.3.B70 Mg2+ required Isodon rubescens
4.2.3.131 Mg2+ required Isodon rubescens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.3.44 (+)-copalyl diphosphate Isodon rubescens
-
 isopimara-7,15-diene + diphosphate
-
 ?
4.2.3.B70 ent-copalyl diphosphate Isodon rubescens
-
 ent-isopimara-7,15-diene + diphosphate
-
 ?
4.2.3.131 (+)-copalyl diphosphate Isodon rubescens
-
 miltiradiene + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.44 Isodon rubescens A0A1Z3GCD1
-
-
4.2.3.B70 Isodon rubescens A0A1X9ISP7
-
-
4.2.3.131 Isodon rubescens A0A1W6QDI7
-
-
4.2.3.185 Isodon rubescens A0A1Z3GBK8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.3.44 recombinant His-tagged KSL6 from Escherichia coli strain Tuner(DE3) by nickel affinity chromatography Isodon rubescens
4.2.3.B70 recombinant His-tagged KSL2 from Escherichia coli strain Tuner(DE3) by nickel affinity chromatography Isodon rubescens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.3.44 leaf
-
 Isodon rubescens
-
4.2.3.B70 leaf
-
 Isodon rubescens
-
4.2.3.131 flower low content of enzyme KSL3, moderate content of enzyme KSL1 Isodon rubescens
-
4.2.3.131 leaf high content of enzyme KSL3, moderate content of enzyme KSL1 Isodon rubescens
-
4.2.3.131 additional information expression patterns of isozymes KSL1 and KSL3, overview Isodon rubescens
-
4.2.3.131 root periderm and cortex, high content of enzyme KSL1, low content of enzyme KSL3 Isodon rubescens
-
4.2.3.131 stem high content of enzyme KSL1, moderate content of enzyme KSL3 Isodon rubescens
-
4.2.3.185 leaf high expression level Isodon rubescens
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.44 (+)-copalyl diphosphate
-
 Isodon rubescens isopimara-7,15-diene + diphosphate
-
 ?
4.2.3.44 additional information GC-MS analysis of compounds Isodon rubescens ?
-
 ?
4.2.3.B70 ent-copalyl diphosphate
-
 Isodon rubescens ent-isopimara-7,15-diene + diphosphate
-
 ?
4.2.3.B70 additional information GC-MS analysis of compounds. The IrKSL2 gene product is an ent-isopimaradiene-like molecule Isodon rubescens ?
-
 ?
4.2.3.131 (+)-copalyl diphosphate
-
 Isodon rubescens miltiradiene + diphosphate
-
 ?
4.2.3.185 ent-copalyl diphosphate
-
 Isodon rubescens ent-atiserene + diphosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.3.44 IrKSL6
-
 Isodon rubescens
4.2.3.44 kaurene synthase 6
-
 Isodon rubescens
4.2.3.44 KSL6
-
 Isodon rubescens
4.2.3.B70 IrKSL2
-
 Isodon rubescens
4.2.3.B70 kaurene synthase 2
-
 Isodon rubescens
4.2.3.B70 KSL2
-
 Isodon rubescens
4.2.3.131 KSL1
-
 Isodon rubescens
4.2.3.131 KSL3
-
 Isodon rubescens
4.2.3.185 kaurene synthase 4
-
 Isodon rubescens
4.2.3.185 KSL4
-
 Isodon rubescens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.3.44 30
-
 assay at Isodon rubescens
4.2.3.B70 30
-
 assay at Isodon rubescens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.3.44 8
-
 assay at Isodon rubescens
4.2.3.B70 8
-
 assay at Isodon rubescens

General Information

EC Number General Information Comment Organism
4.2.3.44 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
4.2.3.44 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
4.2.3.B70 evolution 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 Isodon rubescens
4.2.3.B70 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-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 5.5.1.13) Isodon rubescens
4.2.3.131 metabolism Lamiaceae-specific clade genes (IrCPS1 and IrCPS2) synthesize the intermediate copalyl diphosphate (normal-CPP), while IrCPS4 and IrCPS5 synthesize the intermediate ent-copalyl diphosphate (ent-CPP). IrKSL2, IrKSL4, and IrKSL5 react with ent-CPP to produce an ent-isopimaradiene-like compound, ent-atiserene and ent-kaurene, respectively. Correspondingly, the Lamiaceae-specific clade genes IrKSL1 or IrKSL3 combined with normal-CPP lead to the formation of miltiradiene. Proposed diterpenoid biosynthesis in Isodon rubescens with two sequential diterpene synthases CPS1 and 2 and KSL1 and 3, overview. Miltiradiene is the precursor of abietatriene Isodon rubescens
4.2.3.131 additional information relationship of CPS and KSL genes and metabolic connection Isodon rubescens