德干高原游动放线菌HS-16-20中活性次级代谢产物的分离纯化与结构鉴定

Purification and structure elucidation of bioactive secondary metabolite from Actinoplanes deccanensis HS-16-20

目的德干高原游动放线菌HS-16-20发酵液中活性次级代谢产物的鉴定。方法发酵液经过滤得到菌丝体,菌丝体用乙醇浸泡得乙醇浸泡液。通过HPLC分析与活性测试,确定了浸泡液中具有抑制MRSA活性的峰。浸泡液经过浓缩析出晶体,晶体通过半制备反相高效液相色谱分离得到目标单体化合物,然后利用高分辨质谱和核磁共振等方法确定单体化合物的结构。结果从菌株HS-16-20的发酵液中分离得到1个具有抑制MRSA活性的化合物,通过波谱分析,证实该化合物与台勾霉素B同质。结论从菌株HS-16-20发酵液中分离得到的活性化合物与台勾霉素B同质。文献报道台勾霉素B具有很好的抗MRSA活性,因此台勾霉素B为该菌抗MRSA活性的物质基础。另外,德干高原游动放线菌HS-16-20产台勾霉素B的效价高(>1000mg/L),组分单一,分离纯化工艺简单,非常适合产业化生产。

Objective To identify the active metabolite from the fermentation broth of Actinoplanes deccanensis HS-16-20. Methods The mycelia obtained from the culture broth ofActinoplanes deccanensis HS- 16-20 was extracted with EtOH. By HPLC analysis and activity test, the bioactive peak of the EtOH extract was confirmed. Subsequently, the EtOH extract was evaporated and crystallized to obtain the crystalline compound. The crystalline compound was further purified by semi-preparative HPLC to yield the target metabolite and its structure was elucidated by HR-ESI-MS and NMR analysis. Results A bioactive compound obtained from the fermentation broth of Actinoplanes deccanensis HS-16-20 was identified to be tiacumicin B on the basis of spectral data. Conclusion The bioactive compound isolated from Actinoplanes deccanensis HS-16-20 was elucidated as tiacumicin B. The literature reported that tiacumicin B has good anti-MRSA activity, so tiacumicin B was the material basis for the anti-MRSA activity of the the strain HS-16-20. In addition, the Actinoplanes deccanensis HS-16-20 is a high yield strain of tiacumicin B （〉1000mg/L） and its chemical constituents is single. Thus, the purification process of tiacumicin B developed in this work was simple and very suitable for industrial production.