Synthetic Biology Approaches to New Bisindoles.

Bisindoles are a class of natural products derived from oxidative dimerization of tryptophan, and many of these molecules have potential use as anticancer agents. The recent isolation of new bisindoles and their corresponding gene clusters has greatly expanded the repertoire of biosynthetic genes available to synthetic biologists. This chapter describes methods to exploit the biosynthetic pathways leading to bisindoles, using cladoniamides as a representative example. Specifically, we describe how to identify and heterologously express gene clusters and how to manipulate pathways in order to generate new bisindoles. We also discuss methods for cultivating, extracting, purifying, and characterizing these new metabolites.