该研究采用4种分离培养基对32份土壤样本中的放线菌进行初步分离纯化,通过抗耐甲氧西林金黄色葡萄球菌(MRSA)抑菌活性实验进行菌株初筛,并采用聚合酶链式反应(PCR)扩增菌株16S rRNA和次级代谢相关基因,对活性菌株的代谢潜力进行评估。结果表明,从32份样本中共分离纯化到169株放线菌,其中68株放线菌具有抗MRSA活性。活性菌株主要为链霉菌属(Streptomyces sp.),且16S rRNA基因序列相似性<98%的菌株有10株,卤化酶(Hal)基因阳性菌株占17.65%,聚酮合酶Ⅰ型(PKS Ⅰ)基因阳性菌株占36.47%,聚酮合酶Ⅱ(PKS Ⅱ)基因阳性菌株占61.77%,非核糖体多肽合成酶(NRPS)基因阳性菌株占32.35%。表明黔北地区土样中放线菌资源丰富,具有从中发现放线菌新种和抗生素的潜力。
The Actinomycetes were primary isolated and purified from 32 soil samples using four separation media. The strains were screened out by anti-methicillin-resistant Staphylococcus aureus (MRSA) bacteriostatic test,and the 16S rRNA and secondary metabolism-related genes were amplified by polymerase chain reaction to evaluate the metabolic potential of the active strain. The results showed that 169 strains of Actinomycetes were isolated and purified from 32 samples. Among them,68 strains of Actinomycetes had anti-MRSA activity. The active strains were mainly Streptomyces sp.,and there were 10 strains with 16S rRNA gene sequence similarity less than 98%;halogenase (Hal) gene-positive strains accounted for 17.65%,polyketide synthase type I (PKS I) gene-positive strains accounted for 36.47%,polyketide synthase II (PKS II) gene-positive strains accounted for 61.77%,and non-ribosomal polypeptide synthetase (NRPS) gene-positive strains accounted for 32.35%. It was indicated that the Actinomycetes were rich in soil samples in the northern Guizhou province of China,and had the potential to discover new Actinomycetes and antibiotics with novel structure.
anti-methicillin-resistant Staphylococcus aureus
secondary metabolite products