Literature summary for 4.2.1.133 extracted from

Hansen, N.L.; Nissen, J.N.; Hamberger, B.
Two residues determine the product profile of the class II diterpene synthases TPS14 and TPS21 of Tripterygium wilfordii (2017), Phytochemistry, 138, 52-56 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene TPS21, functional recombinant expression of wild-type and mutant enzymes in Nicotiana benthamiana leaves, and to increase expression level and elevate the GGDP pool, the p19 suppressor of gene silencing and Coleus forskohlii 1-deoxy-D-xylulose 5-phosphate synthase (CfDXS) and GGDP synthase (CfGGDPS) are coexpressed	Tripterygium wilfordii

Protein Variants

Protein Variants	Comment	Organism
A325V	site-directed mutagenesis, the mutant shows substrate specificity of EC 5.5.1.28 in contrast to wild-type	Tripterygium wilfordii
H265Y	site-directed mutagenesis, the mutant shows substrate specificity of EC 5.5.1.28 in contrast to wild-type	Tripterygium wilfordii
H265Y/A325V	site-directed mutagenesis, the mutant shows substrate specificity of EC 5.5.1.28 in contrast to wild-type	Tripterygium wilfordii
additional information	using site-directed mutagenesis, a panel of six variants, where one, or both positions are exchanged between the enzymes TPS14 and TPS21, i.e. kolavenyl diphosphate synthase and copal-8-ol diphosphate hydratase. In coupled heterologous assays with a corresponding class I diTPS, TwTPS2, complete product interchange is observed in variants with both reciprocal mutations, while substitutions of either residue give mixed product profiles. Two mutants, TwTPS14:Y265H and TwTPS21:A325V, also produce ent-copalyl diphosphate, highlighting the evolutionary potential of enzymes of this family to drive rapid diversification of plant diterpene biosynthesis through neo-functionalization	Tripterygium wilfordii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
geranylgeranyl diphosphate + H2O	Tripterygium wilfordii	-	ent-(13E)-8alpha-hydroxylabd-13-en-15-yl diphosphate	-	r

Organism

Organism	UniProt	Comment	Textmining
Tripterygium wilfordii	A0A0M4MZ71	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
geranylgeranyl diphosphate + H2O	-	Tripterygium wilfordii	ent-(13E)-8alpha-hydroxylabd-13-en-15-yl diphosphate	-	r
additional information	coupled heterologous assays with a corresponding class I diTPS, TwTPS2	Tripterygium wilfordii	?	-	?

Synonyms

Synonyms	Comment	Organism
TPS21	-	Tripterygium wilfordii
TwTPS21	-	Tripterygium wilfordii

General Information

General Information	Comment	Organism
evolution	the enzyme is a class II diterpene synthase (diTPS) of specialized labdane-type metabolism. Evolutionary potential of enzymes of this family to drive rapid diversification of plant diterpene biosynthesis through neo-functionalization. Reaction mechanisms of class II diTPSs (a), and labdane-type diterpenes, overview	Tripterygium wilfordii
metabolism	the enzyme is a class II diterpene synthase (diTPS) of specialized labdane-type metabolism	Tripterygium wilfordii
additional information	enzyme structure homology modelling based on an the crystal structure of the Arabidopsis thaliana ent-copalyl diphosphate synthase, EC 5.5.1.13, active structure analysis, structure-function relationship analysis, overview	Tripterygium wilfordii
physiological function	enzyme TwTPS21 catalyzes bicyclization of the linear C20 precursor geranylgeranyl diphosphate to ent-copal-8-ol diphosphate	Tripterygium wilfordii

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Release 2023.1 (January 2023)