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Implication of PKS type Ⅰ gene and chromatographic strategy for the biodiscovery of antimicrobial polyketide metabolites from endosymbiotic Nocardiopsis prasina CLA68

机译:PKSⅠ型基因的涵义和色谱策略对内生共生夜蛾拟南芥CLA68中抗菌聚酮化合物代谢物的生物发现

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摘要

Advanced approach in probing for polyketide antimicrobials requires novel genomics and chromatographic strategies. An endophytic strain CLA68 was isolated from the root of Combretum latifolium Blume (Combretaceae) collected from the Western Ghats of Southern India. Strain CLA68 was then identified as Nocardiopsis prasina by its characteristic culture morphology and analysis of 16S rRNA gene sequence. Biosynthetic polyketide synthase genes were investigated using two pairs of degenerate primers. Ethyl acetate extract of CLA68 exhibited broad spectrum activity against a panel of test human pathogens. PKS type-Ⅰ gene detection and chromatographic strategy yielded a robust polyketide antimicrobial compound which identified as nocapyrone E. Minimum inhibitory concentration of the purified compound against MRSA and other human pathogens ranged between 25 and 100 μg/ml. The present work highlights the utility of N. prasina CLA68 as potential source for antimicrobial polyketide nocapyrone E which could help to combat multidrug-resistant pathogens. This study demonstrates feasibility of PKS type-Ⅰ gene-based molecular approach and chemical investigation by chromatographic approach is the best method for prediction and rapid discovery of novel polyketides from endosymbiotic actinomycetes. The sequence data of this endosymbiotic actinomycete is deposited in GenBank under the accession no. KP269077.
机译:探测聚酮化合物抗菌剂的先进方法需要新颖的基因组学和色谱策略。从印度南部西高止山脉收集的Combretum latifolium Blume(Combretaceae)的根中分离出一种内生菌株CLA68。然后通过其特征性的培养形态和对16S rRNA基因序列的分析,将菌株CLA68鉴定为Pranos。使用两对简并引物研究了生物合成的聚酮化合物合酶基因。 CLA68的乙酸乙酯提取物对一组受测人类病原体表现出广谱活性。通过PKSⅠ型基因检测和色谱分析策略,得到了一种稳健的聚酮化合物抗菌化合物,被鉴定为NocapyroneE。纯化的化合物对MRSA和其他人类病原体的最低抑菌浓度为25至100μg/ ml。目前的工作突出了N. prasina CLA68作为抗菌聚酮Nocapyrone E的潜在来源的实用性,它可以帮助抵抗多药耐药的病原体。本研究证明了基于PKSⅠ型基因的分子方法的可行性,并且通过色谱方法进行化学研究是预测和快速发现内共生放线菌中新型聚酮化合物的最佳方法。该内共生放线菌的序列数据保存在GenBank中,登录号为。 KP269077。

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