   4.2.1.133 | more |
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 |
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