CIRAD, UMR BGPI, Montpellier Cedex 5, F-34398, France.
BMC Genomics. 2013 Sep 27;14:658. doi: 10.1186/1471-2164-14-658.
Various bacteria can use non-ribosomal peptide synthesis (NRPS) to produce peptides or other small molecules. Conserved features within the NRPS machinery allow the type, and sometimes even the structure, of the synthesized polypeptide to be predicted. Thus, bacterial genome mining via in silico analyses of NRPS genes offers an attractive opportunity to uncover new bioactive non-ribosomally synthesized peptides. Xanthomonas is a large genus of Gram-negative bacteria that cause disease in hundreds of plant species. To date, the only known small molecule synthesized by NRPS in this genus is albicidin produced by Xanthomonas albilineans. This study aims to estimate the biosynthetic potential of Xanthomonas spp. by in silico analyses of NRPS genes with unknown function recently identified in the sequenced genomes of X. albilineans and related species of Xanthomonas.
We performed in silico analyses of NRPS genes present in all published genome sequences of Xanthomonas spp., as well as in unpublished draft genome sequences of Xanthomonas oryzae pv. oryzae strain BAI3 and Xanthomonas spp. strain XaS3. These two latter strains, together with X. albilineans strain GPE PC73 and X. oryzae pv. oryzae strains X8-1A and X11-5A, possess novel NRPS gene clusters and share related NRPS-associated genes such as those required for the biosynthesis of non-proteinogenic amino acids or the secretion of peptides. In silico prediction of peptide structures according to NRPS architecture suggests eight different peptides, each specific to its producing strain. Interestingly, these eight peptides cannot be assigned to any known gene cluster or related to known compounds from natural product databases. PCR screening of a collection of 94 plant pathogenic bacteria indicates that these novel NRPS gene clusters are specific to the genus Xanthomonas and are also present in Xanthomonas translucens and X. oryzae pv. oryzicola. Further genome mining revealed other novel NRPS genes specific to X. oryzae pv. oryzicola or Xanthomonas sacchari.
This study revealed the significant potential of the genus Xanthomonas to produce new non-ribosomally synthesized peptides. Interestingly, this biosynthetic potential seems to be specific to strains of Xanthomonas associated with monocotyledonous plants, suggesting a putative involvement of non-ribosomally synthesized peptides in plant-bacteria interactions.
多种细菌可以使用非核糖体肽合成(NRPS)来产生肽或其他小分子。NRPS 机制中的保守特征允许预测合成多肽的类型,有时甚至是结构。因此,通过对 NRPS 基因的计算机分析进行细菌基因组挖掘,为发现新的非核糖体合成的生物活性肽提供了一个有吸引力的机会。黄单胞菌是一个革兰氏阴性细菌的大属,可引起数百种植物物种的疾病。迄今为止,该属中唯一已知的由 NRPS 合成的小分子是由黄单胞菌属 albilineans 产生的 albicidin。本研究旨在通过对最近在测序的黄单胞菌属 albilineans 和相关黄单胞菌属物种的基因组中鉴定出的具有未知功能的 NRPS 基因进行计算机分析,来估计黄单胞菌属的生物合成潜力。
我们对所有已发表的黄单胞菌属基因组序列中的 NRPS 基因以及未发表的水稻白叶枯病菌 pv.oryzae 菌株 BAI3 和黄单胞菌属菌株 XaS3 的草案基因组序列进行了计算机分析。这两个菌株,以及黄单胞菌属 albilineans 菌株 GPE PC73 和水稻白叶枯病菌 pv.oryzae 菌株 X8-1A 和 X11-5A,拥有新的 NRPS 基因簇,并共享相关的 NRPS 相关基因,如非蛋白质氨基酸生物合成或肽分泌所需的基因。根据 NRPS 结构对肽结构的计算机预测表明,每种产生菌株都有八种不同的肽。有趣的是,这八种肽不能分配到任何已知的基因簇中,也与天然产物数据库中的已知化合物无关。对 94 种植物病原细菌的 PCR 筛选表明,这些新的 NRPS 基因簇是黄单胞菌属特有的,也存在于黄单胞菌属 translucens 和水稻白叶枯病菌 pv.oryzicola 中。进一步的基因组挖掘揭示了其他特定于水稻白叶枯病菌 pv.oryzicola 或甘蔗黄单胞菌的新 NRPS 基因。
本研究揭示了黄单胞菌属产生新的非核糖体合成肽的巨大潜力。有趣的是,这种生物合成潜力似乎是与单子叶植物相关的黄单胞菌属菌株所特有的,这表明非核糖体合成的肽可能参与了植物-细菌的相互作用。