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Functional analysis of environmental DNA-derived microviridins provides new insights into the diversity of the tricyclic peptide family

  • Microviridins represent a unique family of ribosomally synthesized cage-like depsipeptides from cyanobacteria with potent protease-inhibitory activities. The natural diversity of these peptides is largely unexplored. Here, we describe two methodologies that were developed to functionally characterize cryptic microviridin gene clusters from metagenomic DNA. Environmental samples were collected and enriched from cyanobacterial freshwater blooms of different geographical origins containing predominantly Microcystis sp. Microviridins were produced either directly from fosmid clones or after insertion of environmental DNA-derived gene cassettes into a minimal expression platform in Escherichia coli. Three novel microviridin variants were isolated and tested against different serine-type proteases. The comparison of the bioactivity profiles of the new congeners allows deduction of further structure-function relationships for microviridins. Moreover, this study provides new insights into microviridin processing and gene cluster organization.Microviridins represent a unique family of ribosomally synthesized cage-like depsipeptides from cyanobacteria with potent protease-inhibitory activities. The natural diversity of these peptides is largely unexplored. Here, we describe two methodologies that were developed to functionally characterize cryptic microviridin gene clusters from metagenomic DNA. Environmental samples were collected and enriched from cyanobacterial freshwater blooms of different geographical origins containing predominantly Microcystis sp. Microviridins were produced either directly from fosmid clones or after insertion of environmental DNA-derived gene cassettes into a minimal expression platform in Escherichia coli. Three novel microviridin variants were isolated and tested against different serine-type proteases. The comparison of the bioactivity profiles of the new congeners allows deduction of further structure-function relationships for microviridins. Moreover, this study provides new insights into microviridin processing and gene cluster organization.show moreshow less

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Metadaten
Author:Douglas Gatte-Picchi, Annika Weiz, Keishi Ishida, Christian Hertweck, Elke Dittmann-ThünemannORCiDGND
DOI:https://doi.org/10.1128/AEM.03502-13
ISSN:0099-2240
ISSN:1098-5336
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=24334668
Parent Title (English):Applied and environmental microbiology
Publisher:American Society for Microbiology
Place of publication:Washington
Document Type:Article
Language:English
Year of first Publication:2014
Year of Completion:2014
Release Date:2017/03/27
Volume:80
Issue:4
Pagenumber:8
First Page:1380
Last Page:1387
Funder:National Council for Scientific and Technological Development (CNPq); German Research Foundation (DFG) [Di910/4-1, He3469/4-1]
Organizational units:Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie
Peer Review:Referiert