基于单菌多次级代谢产物策略的深海来源放线菌Streptomyces sp.SCSIO 15079化学多样性研究

Study on the Chemical Diversity of the Deep-Sea-Derived Actinomycete Streptomyces sp. SCSIO 15079 Based on One-Strain Many Compounds Strategy

关键词:  深海放线菌  OSMAC  发酵方式  化学多样性  次级代谢产物

Key words:  deep-sea actinomycetes  OSMAC  fermentation mode  chemical diversity  secondary metabolites

摘要:

采用单菌多次级代谢产物(OSMAC)策略对1株深海放线菌Streptomyces sp.SCSIO 15079进行化学多样性研究。通过在不同盐度、pH值和发酵时长条件下对菌株进行培养调控并筛选出1种适宜发酵条件，分别进行摇床和发酵罐发酵。综合运用正相硅胶柱色谱、十八烷基硅烷(Octadecyl Silance,ODS)反相柱色谱、葡聚糖凝胶柱色谱(Sephadex LH-20)、半制备高效液相色谱(HPLC)等方法，对菌株摇床与发酵罐发酵产物中具有差异的化学成分进行导向分离与纯化，根据核磁数据和文献对比进行化合物结构鉴定。研究结果表明，将Streptomyces sp.SCSIO 15079菌种接种到不添加海盐、pH值为7.2的A培养基中，于28℃、180 r/min摇床发酵7 d，可获得更丰富的代谢产物；经分离鉴定，从摇床发酵产物中得到的9个与发酵罐发酵产物不一样的化合物且均为含氮类化合物，分别为吲哚甲醛(1)、3-羟基吲哚(2)、胸腺嘧啶(3)、尿嘧啶(4)、环-(脯氨酸-苯丙氨酸)二肽(5)、环-(亮氨酸-缬氨酸)二肽(6)、环-(丙氨酸-异亮氨酸)二肽(7)、环-(丙氨酸-缬氨酸)二肽(8)、环-(脯氨酸-缬氨酸)二肽(9)。上述结果表明改变发酵方式可影响菌株基因的转录和翻译，调控其代谢并产生新的代谢产物。

Abstract:

The chemical diversity of a deep-sea actinomycete Streptomyces sp. SCSIO 15079 was investigated using one strain many compounds (OSMAC) strategy. The strains were cultured and regulated under different salinity,pH value and fermentation time,and one suitable fermentation condition was selected for shaking table and fermentor fermentation,respectively. A combination of normal phase silica gel column chromatography,Octadecyl Silane (ODS),sephadex gel column chromatography (Sephadex LH-20) and semi-preparative High Performance Liquid Chromatography (HPLC) was used to isolate and purify the different chemical components in the fermentation products of the strain shaking table and the fermentor.The structure of the compounds was identified according to the NMR data and literature comparison.The results showed that Streptomyces sp.SCSIO 15079 was inoculated into A medium without sea salt and pH 7.2 and fermented in shaking table at 28℃ and 180 r/min for 7 d to obtain more abundant metabolites. Through separation identification,the nine compounds obtained from the shaking fermentation products that were different from those from the fermentation products in the fermentor were all nitrogenous compounds,they were identified as 1H-indole-3-carbaldehyde (1),3-Hydroxy-1H-indole(2),thymine (3),uiracil (4),Cyclo (D-Phe-L-Pro) (5),Cyclo (Leu-Val) (6),Cyclo (Ala-Val) (7),Cyclo (Ile-Ala) (8),Cyclo (D-Val-L-Pro) (9). It shows that changing the fermentation method can affect the transcription and translation of the strain's genes and regulate its metabolic pathways to produce new metabolites.