A Catalytically Competent Terpene Synthase Inferred Using Ancestral Sequence Reconstruction Strategy
Résumé
On the basis of bioinformatic analysis of 137 putative and characterized bacterial terpene synthases (TS), we applied ancestral reconstruction strategy to design a biosynthetic protein sequence (AncCL1). We biochemically confirmed its catalytic competence and product profile. This enzyme catalyzes not only epi-isozizaene (minor product) but also epi-zizaene (major product) formation. We compared AncCL1 with two related TS to allow a more detailed insight into the product specificity determinants. In the future, the presented method will be useful for engineering biosynthetic TS and to elucidate the function of unknown enzymes.