Linaridins are members of the ribosomally synthesized and post-translationally modified peptide (RiPP) family of natural products. Five linaridins have been reported, which are defined by the presence of dehydrobutyrine, a dehydrated, alkene-containing amino acid derived from threonine. This work describes the development of a linaridin-specific scoring module for Rapid ORF Description and Evaluation Online (RODEO), a genome-mining tool tailored toward RiPP discovery. Upon mining publicly accessible genomes available in the NCBI database, RODEO identified 561 (382 nonredundant) linaridin biosynthetic gene clusters. Linaridin BGCs with unique gene architectures and precursor sequences markedly different from previous predictions were uncovered during these efforts. To aid in data set validation, two new linaridins, pegvadin A and B, were detected through reactivity-based screening and isolated from Streptomyces noursei and Streptomyces auratus, respectively. Reactivity-based screening involves the use of a probe that chemoselectively modifies an organic functional group present in the natural product. The dehydrated amino acids present in linaridins as α/β-unsaturated carbonyls were appropriate electrophiles for nucleophilic 1,4-addition using a thiol-functionalized probe. The data presented within significantly expand the number of predicted linaridin biosynthetic gene clusters and serve as a roadmap for future work in the area. The combination of bioinformatics and reactivity-based screening is a powerful approach to accelerate natural product discovery.

中文翻译：

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基于生物信息学和反应性的 Linaridin 发现

Linaridins 是核糖体合成和翻译后修饰肽 (RiPP) 天然产物家族的成员。已报道了五种里那定，它们是通过存在脱氢丁酸来定义的，脱氢丁酸是一种衍生自苏氨酸的脱水含烯烃氨基酸。这项工作描述了用于快速 ORF 在线描述和评估 (RODEO) 的 linaridin 特异性评分模块的开发，RODEO 是一种针对 RiPP 发现量身定制的基因组挖掘工具。在挖掘 NCBI 数据库中可公开访问的基因组后，RODEO 鉴定了 561 个（382 个非冗余）linaridin 生物合成基因簇。在这些努力中，发现了具有独特基因结构和前体序列的 Linaridin BGC，与之前的预测明显不同。为了帮助数据集验证，通过基于反应性的筛选检测到两种新的linaridins，pegvadin A和B，并分别从Streptomyces noursei和Streptomyces auratus中分离出来。基于反应性的筛选涉及使用化学选择性修饰天然产物中存在的有机官能团的探针。亚油酸中以 α/β-不饱和羰基形式存在的脱水氨基酸是适合使用硫醇官能化探针进行亲核 1,4-加成的亲电子试剂。其中提供的数据显着扩展了预测的里那丁生物合成基因簇的数量，并作为该领域未来工作的路线图。生物信息学和基于反应性的筛选的结合是加速天然产物发现的有效方法。