Functional Analysis of Environmental DNA-Derived Microviridins Provides New Insights into the Diversity of the Tricyclic Peptide Family
Functional Analysis of Environmental DNA-Derived Microviridins Provides New Insights into the Diversity of the Tricyclic Peptide Family
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DOI：
10.1128/aem.03502-13
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发表时间：
2014-02-01
影响因子：
4.4
通讯作者：
Dittmann, Elke
中科院分区：
文献类型：
--
作者：
Gatte-Picchi, Douglas;Weiz, Annika;Dittmann, Elke
Microviridins代表了一个独特的家庭的核糖体合成的笼状缩肽从蓝藻具有强大的蛋白酶抑制活性。这些肽的天然多样性在很大程度上是未开发的。在这里，我们描述了两种方法，开发的功能表征隐蔽microviridin基因簇从宏基因组DNA。从不同地理来源的蓝藻淡水水华中收集和富集环境样品，其中主要含有微囊藻属。微绿素直接从fosmid克隆中产生，或将环境DNA衍生的基因盒插入大肠杆菌的最小表达平台后产生。三种新的microviridin变体被分离并针对不同的丝氨酸型蛋白酶进行测试。新同系物的生物活性谱的比较允许推导microviridins的进一步的结构-功能关系。此外，这项研究提供了新的见解microviridin加工和基因簇组织。
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.