Synthase-Selective Exploration of a Tunicate Microbiome by Activity-Guided Single-Cell Genomics
| Autor: | Katherine Charov, James J. La Clair, Ramunas Stepanauskas, Eric D. Becraft, Frederik Schulz, Woojoo E. Kim, Michael D. Burkart, Mária Džunková, Tanja Woyke, Julia M. Brown |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Secondary Metabolism Siderophores Genomics Computational biology 01 natural sciences Biochemistry Oceanospirillales Article 03 medical and health sciences Polyketide Nonribosomal peptide Gene cluster Genetics Escherichia coli 2.1 Biological and endogenous factors Animals Microbiome Aetiology Peptide Synthases Gene Phylogeny chemistry.chemical_classification biology 010405 organic chemistry Microbiota Human Genome Organic Chemistry Computational Biology General Medicine Cell sorting Biological Sciences biology.organism_classification Flow Cytometry 0104 chemical sciences Ciona intestinalis 030104 developmental biology chemistry Multigene Family Polyketides Chemical Sciences Molecular Medicine Single-Cell Analysis Carrier Proteins Biotechnology Bacillus subtilis |
| Zdroj: | ACS Chemical Biology . ACS Chem Biol ACS chemical biology, vol 16, iss 5 |
| Popis: | While thousands of environmental metagenomes have been mined for the presence of novel biosynthetic gene clusters, such computational predictions do not provide evidence of their in vivo biosynthetic functionality. Using a fluorescent in situ enzyme assay targeting carrier proteins common to polyketide (PKS) and non–ribosomal peptide synthetases (NRPS), we applied fluorescence–activated cell sorting to tunicate microbiome to enrich for microbes with active secondary metabolic capabilities. Single–cell genomics uncovered the genetic basis for a wide biosynthetic diversity in the enzyme–active cells and revealed a member of marine Oceanospirillales harboring a novel NRPS gene cluster with high similarity to phylogenetically distant marine and terrestrial bacteria. Interestingly, this synthase belongs to a larger class of siderophore biosynthetic gene clusters commonly associated with pestilence and disease. This demonstrates activity–guided single–cell genomics as a tool to guide novel biosynthetic discovery. |
| Databáze: | OpenAIRE |
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