Ma Liang, Zhang Wenjun, Zhu Yiguang, Wu Zhengchao, Saurav Kumar, Hang Hui, Zhang Changsheng
CAS Key Laboratory of Tropical Marine Bio-resource and Ecology, RNAM Center for Marine Microbiology, Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China.
Wei Sheng Wu Xue Bao. 2013 Oct 4;53(10):1063-71.
The present study aims to isolate and identify actinobacteria associated with the soft coral Nephthea sp., and to isolate natural products from these actinobacteria under the guidance of PCR screening for polyketides synthase (PKS) genes.
Eleven selective media were used to isolate actinobacteria associated with the soft coral Nephthea sp. collected from Yongxin Island. The isolated actinobacteria were classified on the basis of phylogenetic tree analysis of their 16S rRNA genes. Degenerated primers targeted on conserved KS (ketoacyl-synthase) domain of type I PKS genes were used to screen for potential isolates. The positive isolates were cultured in three different media to check their producing profiles. One bioactive strain that is rich in metabolites was subjected to larger scale fermentation for isolating bioactive natural products.
A total of 20 strains were isolated from Nephthea sp., and were categorized into 3 genera including Streptomyces, Dietzia and Salinospora, among which 18 strains were positive in screening with type I PKS genes. Two bioactive compounds rifamycin S and rifamycin W were isolated and identified from Salinospora arenicola SH04.
This is the first report of isolating indigenous marine actinobacteria Salinospora from the soft coral Nephthea sp. It provides an example of isolating bioactive secondary metabolites from cultivable actinobacteria associated with Nephthea sp. by PCR screening.
本研究旨在分离和鉴定与软珊瑚肾形珊瑚属相关的放线菌,并在聚酮合酶(PKS)基因的PCR筛选指导下,从这些放线菌中分离天然产物。
使用11种选择性培养基分离从永兴岛采集的与软珊瑚肾形珊瑚属相关的放线菌。根据其16S rRNA基因的系统发育树分析对分离出的放线菌进行分类。使用针对I型PKS基因保守KS(酮酰基合酶)结构域的简并引物筛选潜在的分离株。将阳性分离株在三种不同的培养基中培养以检查其生产概况。对一种富含代谢产物的生物活性菌株进行大规模发酵,以分离生物活性天然产物。
从肾形珊瑚属中共分离出20株菌株,分为链霉菌属、迪茨氏菌属和盐孢菌属3个属,其中18株在I型PKS基因筛选中呈阳性。从沙栖盐孢菌SH04中分离并鉴定出两种生物活性化合物利福霉素S和利福霉素W。
这是首次从软珊瑚肾形珊瑚属中分离出土著海洋放线菌盐孢菌属的报道。它提供了一个通过PCR筛选从与肾形珊瑚属相关的可培养放线菌中分离生物活性次生代谢产物的实例。