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16S rRNA phylogenetic analysis of actinomycetes isolated from Eastern Ghats and marine mangrove associated with antibacterial and anticancerous activities

机译:从东高止山脉和海洋红树林中分离的放线菌与抗菌和抗癌活性相关的16S rRNA系统发育分析

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Morphologically 238 different marine and terrestrial actinomycete strains were chosen by screening of the antagonistic activity using human pathogens and cancerous cell lines, for which 185 sediments/rhizosphere soil samples were collected from the different sites of Manakkudi estuary of south-west coastal regions and Yercaud hills of eastern Ghats of Tamil Nadu, India. The present study?aimed?at the evaluation of biologically diverse strains of actinomycetes?from soil samples, for the production of wide spectrum of bioactive secondary metabolites. Most of the isolated strains belong to the genus namely?Streptomyces?(68%), followed by?Actinobacterium?(28%)?andNocardiopsis?(4%). Among the different strains purified, about 10 strains exhibited high antagonistic activity. An attempt was made to study the bioactive compounds extracted from the Act2 strain isolated from marine mangrove sediments, which showed the highest antibacterial and anticancerous activities. The partially purified fifth active fraction was analyzed by Gas chromatography-mass spectrometry (GC-MS) and the possible closely related compounds were identified as Silane, Pyridine, 2,4,6-trimethyl, Amino malonic acid, 4-benzoxazin, Tris methyl and Cyclohexydimethoxy methyl compounds. Studies were conducted to extract the genomic DNA from the selected strains, amplified through polymerase chain reaction (PCR) and subjected to 16S rRNA sequence analysis. Subsequently, phylogenetic tree was constructed using suitable bioinformatics tools to identify the similarity which showed?97%?similarity between strains. Moreover, all the selected strains of actinomycetes were subjected to study the protein and plasmid DNA expression profiles which showed prominent bands with genetically excellent relationship between strains in the profiles.
机译:通过使用人类病原体和癌细胞系筛选拮抗活性,从形态上选择了238种不同的海洋和陆地放线菌菌株,从西南沿海地区Manakkudi河口和Yercaud丘陵的不同地点收集了185个沉积物/根际土壤样品印度泰米尔纳德邦的东高止山脉。本研究的目的是评估土壤样品中放线菌的多种生物菌株,以生产各种生物活性次生代谢产物。分离出的大多数菌株属于链霉菌属(68%),其次是放线杆菌(28%)和诺卡氏菌(4%)。在纯化的不同菌株中,约10个菌株表现出高拮抗活性。试图研究从海洋红树林沉积物中分离的Act2菌株提取的生物活性化合物,该菌株显示出最高的抗菌和抗癌活性。通过气相色谱-质谱(GC-MS)分析部分纯化的第五种活性成分,并鉴定出可能密切相关的化合物为硅烷,吡啶,2,4,6-三甲基,氨基丙二酸,4-苯并嗪,Tris甲基和环己二甲氧基甲基化合物。进行了研究以从选定的菌株中提取基因组DNA,通过聚合酶链反应(PCR)进行扩增,并进行16S rRNA序列分析。随后,使用合适的生物信息学工具构建了系统进化树,以鉴定出显示菌株之间“ 97%”相似性的相似性。此外,对所有选择的放线菌菌株进行了蛋白质和质粒DNA表​​达图谱的研究,这些图谱显示了突出的条带,在图谱中的菌株之间具有遗传学上的极好的关系。

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