为寻找生物活性肽代谢产物而对土壤放线菌分离株进行分子、化学和生物学筛选。
Molecular, chemical and biological screening of soil actinomycete isolates in seeking bioactive peptide metabolites.
作者信息
Hamedi Javad, Imanparast Somaye, Mohammadipanah Fatemeh
机构信息
Department of Microbial Biotechnology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, 14155-6455, Tehran, Iran. ; Microbial Technology and Products Research Center, University of Tehran, Tehran, Iran.
出版信息
Iran J Microbiol. 2015 Feb;7(1):23-30.
BACKGROUND AND OBJECTIVE
Due to the evolution of multidrug-resistant strains, screening of natural resources, especially actinomycetes, for new therapeutic agents discovery has become the interests of researchers. In this study, molecular, chemical and biological screening of soil actinomycetes was carried out in order to search for peptide-producing actinomycetes.
MATERIALS AND METHODS
60 actinomycetes were isolated from soils of Iran. The isolates were subjected to molecular screening for detection NRPS (non-ribosomal peptide synthetases) gene. Phylogenic identification of NRPS containing isolates was performed. Chemical screening of the crude extracts was performed using chlorine o-dianisidine as peptide detector reagent and bioactivity of peptide producing strains was determined by antimicrobial bioassay. High pressure liquid chromatography- mass spectrometry (HPLC-MS) with UV-visible spectroscopy was performed for detection of the metabolite diversity in selected strain.
RESULTS
Amplified NRPS adenylation gene (700 bp) was detected among 30 strains. Phylogenic identification of these isolates showed presence of rare actinomycetes genera among the isolates and 10 out of 30 strains were subjected to chemical screening. Nocardia sp. UTMC 751 showed antimicrobial activity against bacterial and fungal test pathogens. HPLC-MS and UV-visible spectroscopy results from the crude extract showed that this strain has probably the ability to produce new metabolites.
CONCLUSION
By application of a combined approach, including molecular, chemical and bioactivity analysis, a promising strain of Nocardia sp. UTMC 751 was obtained. This strain had significant activity against Staphylococcus aureus and Pseudomonas aeruginosa. Strain Nocardia sp. UTMC 751 produce five unknown and most probably new metabolites with molecular weights of 274.2, 390.3, 415.3, 598.4 and 772.5. This strain had showed 99% similarity to Nocardia ignorata DSM 44496 T.
背景与目的
由于多重耐药菌株的演变，筛选自然资源，尤其是放线菌，以发现新的治疗药物已成为研究人员的兴趣所在。在本研究中，对土壤放线菌进行了分子、化学和生物学筛选，以寻找产生肽的放线菌。
材料与方法
从伊朗土壤中分离出60株放线菌。对分离株进行分子筛选以检测NRPS（非核糖体肽合成酶）基因。对含有NRPS的分离株进行系统发育鉴定。使用氯邻联茴香胺作为肽检测试剂对粗提物进行化学筛选，并通过抗菌生物测定法测定产生肽的菌株的生物活性。采用高效液相色谱 - 质谱联用（HPLC-MS）和紫外可见光谱法检测所选菌株中的代谢物多样性。
结果
在30株菌株中检测到扩增的NRPS腺苷化基因（700 bp）。对这些分离株的系统发育鉴定表明，分离株中存在罕见的放线菌属，30株中有10株进行了化学筛选。诺卡氏菌属UTMC 751对细菌和真菌测试病原体显示出抗菌活性。粗提物的HPLC-MS和紫外可见光谱结果表明，该菌株可能具有产生新代谢物的能力。
结论
通过应用包括分子、化学和生物活性分析在内的联合方法，获得了一株有前景的诺卡氏菌属UTMC 751。该菌株对金黄色葡萄球菌和铜绿假单胞菌具有显著活性。诺卡氏菌属UTMC 751菌株产生了五种分子量分别为274.2、390.3、415.3、598.4和772.5的未知且很可能是新的代谢物。该菌株与无名诺卡氏菌DSM 44496 T显示出99%的相似性。
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