For decades, fungi have been an important source of medically relevant natural products (NPs). Recent advances in DNA sequencing have revealed that the biosynthetic potential of fungal genomes is much deeper than previously realized. Difficulties in culturing and genetically engineering many fungi, combined with the fact that many NP biosynthetic gene clusters (BGCs) are not expressed under laboratory conditions has lead to much of this biosynthetic potential remaining untapped. Here we describe the realization of a pipeline for the high-throughput characterization of fungal NP biosynthetic gene clusters. Our pipeline encompasses: * bioinformatic tools for identification of BGCs and sequence editing (e.g. intron removal) for improved expression in S. cerevisiae * an assembly platform that combines commercial gene synthesis and yeast homologous recombination for the cost-effective construction of large (&gt; 20 kb) DNAs * high-throughput untargeted LC-MS based metabolomics screening of recombinant yeast cultures for the rapid detection and prioritization of potential novel compounds With this pipeline, we successfully detected potentially novel NPs from several previously unstudied fungal BGCs, and have structurally characterized a subset of the BGC-associated compounds. Our pipeline shows how high-throughput synthetic biology tools can facilitate the rapid discovery and biosynthetic production of complex chemical scaffolds of potential pharmaceutical relevance.

Bio-based products: Synthetic biology approaches to engineer production of biofuels & value-added chemicals

253rd National Meeting (2017)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.