Assembly and characterization fungal biosynthetic gene clusters in the yeast Saccharomyces cerevisiae

For decades, fungi have been a vital source of therapeutically relevant natural products. The increase in the number of available genomic and metagenomic datasets over the past 15 years has revealed that the biosynthetic potential of many fungal strains is much greater than previously realized, with the majority of biosynthetic gene clusters (BGCs) remaining silent under standard laboratory culture conditions.

Owing to the difficulties associated with culturing and engineering many fungal species, heterologous expression of these silent BGCs in model organisms is among the most promising means of accessing this untapped biosynthetic diversity.

Here, we present a suite of tools for the high-throughput discovery, assembly, and analysis of fungal BGCs in the model fungus Saccharomyces cerevisiae. Our pipeline principally consists of bioinformatic tools for BGC discovery, a platform for the assembly and sequence verification of large, multi-gene, expression constructs; and high-throughput untargeted metabolomics for the detection of dereplication of compounds produced from recombinant yeast cultures.