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5.5.1.12: copalyl diphosphate synthase

This is an abbreviated version!
For detailed information about copalyl diphosphate synthase, go to the full flat file.

Word Map on EC 5.5.1.12

Reaction

geranylgeranyl diphosphate
=
(+)-copalyl diphosphate

Synonyms

(+)-copalyl diphosphate synthase, (+)-CPS, (-)-abietadiene synthase, abietadiene cyclase, abietadiene synthase, AGAS, BDTC, bifunctional diterpene cyclase, bifunctional LAS-type diTPS, class II diterpene synthase, class II diTPS, CPP synthase, CPS, CPS1, CPS2, CPS3, cyclase, abietadiene, diTPS-ISO, ISO, isopimaradiene synthase-type diTPS, LAS, LAS1, levopimaradiene/abietadiene synthase-type diTPS, More, MvCPS3, PaLAS, RoCPS1, SmCPS, SmCPS1, SmCPS2, TPS-LAS, TPS2

ECTree

     5 Isomerases
         5.5 Intramolecular lyases
             5.5.1 Intramolecular lyases (only sub-subclass identified to date)
                5.5.1.12 copalyl diphosphate synthase

Cloned

Cloned on EC 5.5.1.12 - copalyl diphosphate synthase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis of 18 different amino acid sequence of copalyl diphosphate synthase, CPS, phylogenetic analysis and tree
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expression in Escherichia coli
expression of His-tagged enzyme in Escherichia coli
functional expression of Picea abies isopimara-7,15-diene synthase (diTPS-ISO) in Nicotiana benthamiana leaves via Agrobacterium tumefaciens transfection method, coexpression with a multifunctional cytochrome P450 (CYP720B4) from Picea sitchensis, subcloning in Escherichia coli strain NEB10beta
gene CPS1, phylogenetic analysis and tree, functional recombinant expression in Saccharomyces cerevisiae strain AM113, with co-overexpressing of GGDPS1 from Cistus creticus, and Nicotiana benthamiana, heterologous expression of the RoCPS1 protein in Escherichia coli fails
gene CPS3, phylogenetic analysis and tree, quantitative expression analysis, functional expression of MvCPS3 alone or with MvELS (multifunctional betaalpha-domain class I diterpene synthase) or MvEKS (ent-kaurene synthase) in engineered Nicotiana benthamiana
gene SmCPS1, phylogenetic analysis, a 289-bp gene-specific sequence including the 3' UTR of SmCPS1 is amplified by PCR using cDNA as a template and then cloned into the pK7GWIWG (II) binary, then introduced into Agrobacterium tumefaciens strain EHA105 by electroporation for ransformation of leaf explants from Salvia miltiorrhiza bh2-7 plants, recombinant expression of His-tagged enzyme in Escherichia coli strains Tuner (DE3) or Origami B (DE3)
A0A0A7AN31, A0A0A7ANT1
gene SmCPS2, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strains Tuner (DE3) or Origami B (DE3)
A0A0A7AN31, A0A0A7ANT1
gene TPS-LAS, recombinant expression in Saccharomyces cerevisiae strain AM94, a yeast strain engineered for the production of isoprenoids, which contains three chromosomally integrated copies of a degradation stabilized variant of HMG2 and a heterozygous deletion of ERG9, and in strain AM204, each with or without coexpression of yeast protein Erg20p(F96C), a yeast FPP synthase, overview
Site-directed mutagenesis is carried out via PCR amplification with overlapping mutagenic primers, and the mutant genes verify by complete sequencing. The resulting wild type and mutant genes are then transferred via directional recombination to the T7-promoter and N-terminal 6xhis fusion expression vector pDEST17. Use of the pDEST17 vector results in a 25 amino acid residue linker between the 6 His-tag and the cloned protein.
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