High-Throughput Screening for Terpene-Synthase-Cyclization Activity and Directed Evolution of a Terpene Synthase

A synthetic substrate enables a new colorimetric screen for terpene synthase cyclization activity, facilitating the engineering of these enzymes. Using directed evolution, the thermostability of the sesquiterpene synthase BcBOT2 was increased without the loss of other properties. The technique also enabled rapid optimization of conditions for expression and stabilization in lysate of another terpene synthase, SSCG_02150.