Xiong Zhi-Qiang, Liu Qiao-Xia, Pan Zhao-Long, Zhao Na, Feng Zhi-Xiang, Wang Yong
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
Arch Microbiol. 2015 Mar;197(2):299-309. doi: 10.1007/s00203-014-1059-y. Epub 2014 Nov 23.
Marine actinomycetes are a potential source of a wide variety of bioactive natural products. In this work, seven pretreatments, three selective isolation media, and five artificial seawater concentrations were used to isolate actinomycetes from the sediments collected from Yellow Sea, China. Statistical analysis showed that only the isolation medium strongly affected the total and bioactive numbers of actinomycete isolates. A total of 613 actinobacterial strains were isolated and screened for antimicrobial activities; 154 isolates showed activity against at least one of nine test drug-resistant microorganisms. Eighty-nine representatives with strong antimicrobial activity were identified phylogenetically based on 16S rRNA gene sequencing, which were assigned to five different actinomycete genera Streptomyces, Kocuria, Saccharomonospora, Micromonospora, and Nocardiopsis. Using PCR-based screening for six biosynthetic genes of secondary metabolites, all 45 isolates with acute activity have at least one biosynthetic gene, 28.8 % of which possess more than three biosynthetic genes. As a case, strain SMA-1 was selected for antimicrobial natural product discovery. Three diketopiperazine dimers including a new compound iso-naseseazine B (1) and two known compounds naseseazine B (2) and aspergilazine A (3) were isolated by bioassay-guided separation. These results suggested that actinomycetes from marine sediments are a potential resource of novel secondary metabolites and drugs.
海洋放线菌是多种生物活性天然产物的潜在来源。在本研究中,采用了七种预处理方法、三种选择性分离培养基和五种人工海水浓度,从中国黄海采集的沉积物中分离放线菌。统计分析表明,只有分离培养基对放线菌分离物的总数和生物活性数量有强烈影响。共分离出613株放线菌菌株,并筛选其抗菌活性;154株分离物对九种受试耐药微生物中的至少一种有活性。基于16S rRNA基因测序,对89株具有强抗菌活性的代表菌株进行了系统发育鉴定,它们被归为五个不同的放线菌属,即链霉菌属、考克氏菌属、糖单孢菌属、小单孢菌属和诺卡氏放线菌属。通过基于PCR的方法筛选次生代谢物的六个生物合成基因,所有45株具有活性的分离物都至少有一个生物合成基因,其中28.8%拥有三个以上的生物合成基因。作为一个实例,选择菌株SMA-1来发现抗菌天然产物。通过生物活性导向分离,分离出三种二酮哌嗪二聚体,包括一种新化合物异纳塞嗪B(1)和两种已知化合物纳塞嗪B(2)和曲霉嗪A(3)。这些结果表明,海洋沉积物中的放线菌是新型次生代谢物和药物的潜在资源。