Terpenoids are the largest class of natural products and have played an outsized role in the evolution and adaptation of living systems. Despite the well-documented functional versatility of terpenoids, the evolutionary processes that allow them to address new biochemical challenges are poorly understood and difficult to recapitulate in engineered systems. In this study, we modified <i>Escherichia coli</i> with a synthetic objective—the inhibition of protein tyrosine phosphatase 1B (PTP1B) from <i>Homo sapiens</i>—and used it to evolve terpene synthases (TSs) that achieve this objective. PTP1B is an influential regulatory enzyme, an important model system for biophysical studies, and an elusive drug target; new inhibitors could find broad use. Using our artificial objective as a guide, we uncovered TS mutants in which mutations confer a survival advantage by enabling inhibitor biosynthesis, improving TS solubility, and/or depleting FPP, a toxic intermediate. The best performing mutants exhibited distinct product profiles but included a shared major product that inhibits PTP1B—an example of convergent evolution. These mutants show how TSs can evolve with heterologous hosts to build molecules that solve new biochemical challenges. We will will also discuss approaches for using next-generation sequencing (NGS) to expand the biosynthetic search space.

Evolution-guided biosynthesis of biologically active terpenoids

ACS Spring 2022

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