Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.
World J Microbiol Biotechnol. 2012 May;28(5):2125-37. doi: 10.1007/s11274-012-1018-1. Epub 2012 Feb 15.
The present study aimed to isolate actinobacteria from soil samples and characterized them using molecular tools and screened their secondary metabolites for antimicrobial activities. Thirty-nine strains from four different location of Barrientos Island, Antarctica using 12 types of isolation media was isolated. The isolates were preceded to screening of secondary metabolites for antimicrobial and antifungal activities. Using high-throughput screening methods, 38% (15/39) of isolates produced bioactive metabolites. Approximately 18% (7/39), 18% (7/39), 10% (4/39) and 2.5% (1/39) of isolates inhibited growth of Candida albicans ATCC 10231(T), Staphylococcus aurues ATCC 51650(T), methicillin-resistant Staphylococcus aurues (MRSA) ATCC BAA-44(T) and Pseudomonas aeruginosa ATCC 10145(T), respectively. Molecular characterization techniques like 16S rRNA analysis, Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), Random amplified polymorphic DNA (RAPD) and composite analyses were used to characterize the actinobacteria strains. Analysis of 16S rRNA sequences is still one of the most powerful methods to determine higher taxonomic relationships of Actinobacteria. Both RAPD and ERIC-PCR fingerprinting have shown good discriminatory capability but RAPD proved to be better in discriminatory power than ERIC-PCR. Our results demonstrated that composite analysis of both fingerprinting generally increased the discrimination ability and generated best clustering for actinobacteria strains in this study.
本研究旨在从土壤样本中分离放线菌，并用分子工具对其进行表征，并筛选其次级代谢产物的抗菌活性。使用 12 种分离培养基，从南极洲巴伦蒂奥斯岛的四个不同地点分离出 39 株菌。将这些分离株进行次级代谢产物的抗菌和抗真菌活性筛选。使用高通量筛选方法，38%（15/39）的分离株产生了具有生物活性的代谢产物。大约 18%（7/39）、18%（7/39）、10%（4/39）和 2.5%（1/39）的分离株分别抑制了白色念珠菌 ATCC 10231（T）、金黄色葡萄球菌 ATCC 51650（T）、耐甲氧西林金黄色葡萄球菌（MRSA）ATCC BAA-44（T）和铜绿假单胞菌 ATCC 10145（T）的生长。使用 16S rRNA 分析、肠细菌重复基因间一致性聚合酶链反应（ERIC-PCR）、随机扩增多态性 DNA（RAPD）和综合分析等分子特征技术对放线菌菌株进行了表征。16S rRNA 序列分析仍然是确定放线菌高级分类关系最有力的方法之一。RAPD 和 ERIC-PCR 指纹图谱都显示出良好的区分能力，但 RAPD 比 ERIC-PCR 具有更好的区分能力。我们的结果表明，两种指纹图谱的综合分析通常可以提高对放线菌菌株的区分能力，并在本研究中产生最佳聚类。
