海绵共附生放线菌Streptomyces parvulus MA1076的次级代谢产物研究

Secondary metabolites from a marine sponge-derived actinomycete Streptomyces parvulus MA1076

海绵共附生放线菌是活性次级代谢产物的重要来源,对海绵共附生放线菌Streptomyces parvulus MA1076进行菌株鉴定和化学成分分析。通过16S rDNA序列分析、构建系统发育树进行菌株鉴定;利用硅胶柱色谱、Sephadex LH-20凝胶柱色谱以及半制备高效液相色谱等技术对其发酵产物进行分离和纯化,通过核磁共振、高分辨质谱等方法,并结合文献数据对化合物进行结构鉴定。从放线菌Streptomyces parvulus MA1076中分离鉴定出6个化合物,分别为放线菌素D(化合物1)、胡萝卜苷(化合物2)、环-((S)-脯氨酸-(R)-亮氨酸)(化合物3)、胸腺嘧啶核苷(化合物4)、3′-氧-乙酰胸腺嘧啶核苷(化合物5)和3-羧基吲哚(化合物6)。化合物2~6为首次从该种属放线菌中分离得到。活性筛选结果显示,化合物1对两株胰腺癌细胞系有明显的增殖抑制活性,研究结果丰富了海绵共附生放线菌次级代谢产物的结构多样性,可为开发利用海洋微生物资源提供参考。

In recent years,researchers have discovered a large number of secondary metabolites with novel chemical structures and diverse activities from sponge-derived actinomycetes.Sponge-derived actinomycetes have become important sources of new drug lead compounds.The actinomycete Streptomyces parvulus MA1076 is derived from the sponge(Pseudoceratina sp.)on Yongxing Island in the South China Sea.In order to obtain the structural type and potential medicinal value of the secondary metabolites,strain identification and chemical composition analysis were carried out on the sponge-associated actinomycete Streptomyces parvulus MA1076.Bacterial genomic DNA rapid extraction kit was used to extract streptomyces genomic DNA.Streptomyces universal primers 27F and 1492R were used for PCR amplification,and the amplified products was sent for sequence testing.The sequencing result was used for strain identification through 16S rDNA sequence analysis and phylogenetic tree construction.Silica gel column chromatography,Sephadex LH-20 gel column chromatography,semi-preparative high performance liquid chromatography and other techniques were applied to separate and purify the ethyl acetate extract of the sponge-derived actinomycetes Streptomyces parvulus MA1076.The structure of the compound was identified by spectral data such as proton nuclear magnetic resonance spectroscopy(1 H-NMR),carbon nuclear magnetic resonance spectroscopy(13 C-NMR),high-resolution mass spectrometry(HR-MS),and combined with the comparison with literature data.Six compounds were isolated and identified from the actinomycetes Streptomyces parvulus MA1076,which were actinomycin D(compound 1),daucosterol(compound 2),cyclo-((S)-Pro-(R)-Leu)(compound 3),thymidine(compound 4),3′-O-acetylthymidine(compound 5),and indole-3-carboxyli acid(compound 6).Compounds 2-6 were isolated for the first time from Streptomyces parvulus MA1076.The six compounds were tested for their cell proliferation inhibitory activity on two pancreatic cancer cell lines(pc-12 and bc-16).Results showed that compound 1 had obvious proliferation inhibitory activity on both pancreatic cancer cell lines.Compounds 2-6 showed no obvious anti-tumor cell proliferation activity.The results of the study make full use of the abundant Chinese sponge resources and their associated microbial resources,exploring the structural diversity of sponge secondary metabolites to discover new chemical substances that may be contained in natural products,and to develop new antibacterial,antitumor drugs is of great theoretical significance and potential application value.