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Targeted Metagenomics: Finding Rare Tryptophan Dimer Natural Products in the Environment

Author:
Chang, Fang-Yuan, Ternei, Melinda A., Calle, Paula Y., Brady, Sean F.
Source:
Journal of the American Chemical Society 2015 v.137 no.18 pp. 6044-6052
ISSN:
1520-5126
Subject:
DNA, bioactive properties, genetic variation, genomic libraries, heterologous gene expression, metagenomics, multigene family, nucleotide sequences, phylogeny, screening, surveys, tryptophan
Abstract:
Natural product discovery from environmental genomes (metagenomics) has largely been limited to the screening of existing environmental DNA (eDNA) libraries. Here, we have coupled a chemical-biogeographic survey of chromopyrrolic acid synthase (CPAS) gene diversity with targeted eDNA library production to more efficiently access rare tryptophan dimer (TD) biosynthetic gene clusters. A combination of traditional and synthetic biology-based heterologous expression efforts using eDNA-derived gene clusters led to the production of hydroxysporine (1) and reductasporine (2), two bioactive TDs. As suggested by our phylogenetic analysis of CPAS genes, identified in our survey of crude eDNA extracts, reductasporine (2) contains an unprecedented TD core structure: a pyrrolinium indolocarbazole core that is likely key to its unusual bioactivity profile. This work demonstrates the potential for the discovery of structurally rare and biologically interesting natural products using targeted metagenomics, where environmental samples are prescreened to identify the most phylogenetically unique gene sequences and molecules associated with these genes are accessed through targeted metagenomic library construction and heterologous expression.
Agid:
5410049