Harnessing the diversity of ATP grasp-type depsipeptide biosyntheses in cyanobacteria: genomic mining and engineering

上个世纪基于天然产物的药物开发主要集中在四类化合物：萜类化合物、生物碱、聚酮化合物和非核糖体肽。在过去的几年里，另一组代谢物，即核糖体产生和翻译后修饰的肽（RiPP）引起了越来越多的关注。我们最近已经显示出巨大的工程潜力的microviridin针对不同的丝氨酸型蛋白酶的优化。然而，该系统的局限性在于肽的环大小的低混杂性。因此，在本项目中，我们的目标是分析进一步的ATPgrasp型RiPP生物合成蓝藻，我们预计替代环的大小。具体来说，我们打算通过使用不同的基因组挖掘策略来识别肽。我们进一步的目的是了解决定环的大小和前导肽的具体作用的机制基础。我们将评估至少一个新的ATPgrasp型RiPP的合理和随机的工程方法的潜力。最后，我们将评估新肽的生物活性，并计划通过噬菌体展示开发高通量筛选系统。该研究的首要目标是开发一个平台，用于定制设计不同环大小的环状缩酚肽，这些缩酚肽可以针对不同的靶标进行筛选。

Natural product-based drug development in the last century was largely focussing on four groups of compounds: terpenoids, alkaloids, polyketides and non-ribosomal peptides. In the past few years, another group of metabolites, ribosomally produced and posttranslationally modified peptides (RiPPs) attracted increasing attention. We have recently shown the great engineering potential of microviridin for the optimization against different serine type proteases. The limitation of the system, however, is the low promiscuity in the ring size of the peptides. In the present project, we therefore aim to analyze further ATPgrasp type RiPP biosyntheses from cyanobacteria for which we anticipate alternative ring sizes. Specifically, we intend to identify the peptides by using different genomic mining strategies. We further aim to understand the mechanistic basis that determines the ring size and the specific role of the leader peptide. We will evaluate the potential of at least one novel ATPgrasp type RiPP for rational and random engineering approaches. Finally, we will assess the bioactivity of the novel peptides and plan to develop a high throughput screening system via phage display. The overarching aim of the study is to develop a platform for the tailor-made design of cyclic depsipeptides of different ring sizes that can potentially be screened against diverse targets.