拟黑多刺蚁共生放线菌的种群组成及其代谢新活性化合物研究

Microbial natural products have been the major source for lead compounds. As special environmental microorganisms, insect symbiotic actinobacteria have yielded many compounds with novel structures, and their continued mining holds much promise for the discovery of microbial drug. Polyrhachis vicina Roger is the only ant that has been approved as food and drug by the Ministry of Health of the People’s Republic of China. It has several cure and healthcare functions, e.g. anti-inflammation, anticancer and regulating immune system, which may be related to its symbiotic microorgnisms because of the long time coevolution. However, up to now, there are not any reseaches about Polyrhachis vicina Roger symbiotic microorgnisms. During the investigation of the symbiotic association between actinobacteria and Polyrhachis vicina Roger, 325 symbiotic actinobacteria was isolated，and 1 novel comound with antitumor and antifungal activity was identified. Subsequently, we explored symbiotic microbial diversity and taxonomy of one sample using 16S rRNA hypervariable tag sequencing, the result showed that Actinobacteria was the second largest phylum symbioting with Polyrhachis vicina Roger, which contains lots of Actinomycetaceae. To explore specific symbiotic actinobacteria associated with Polyrhachis vicina Roger, we perform 16S rRNA hypervariable tag sequencing on multiple samples in this project. Special media or separation methods are then employed to isolate them. Then a rapid and efficient method (LC/MS-Principal Component Analysis) is employed to screen novel compounds from them.Subsequently, the novel compounds are isolated and characterized, and their antibacterial, antifungal, insecticidal and antitumor activities are evaluted, looking forward to obtaining bioactive compounds with novel structures or new role characteristics.

微生物天然产物一直是人类获得先导化合物的重要来源。昆虫共生放线菌作为一类特境微生物，合成多种新奇结构化合物，为发现新的微生物天然产物药物提供可能。拟黑多刺蚁是我国卫生部唯一批准的药食两用蚁，有消炎、抗肿瘤及增强免疫力等功效，其许多功效可能与其共生微生物有关，但目前关于其共生微生物的研究未见报道。申请者在对其共生放线菌的研究中，分离放线菌325株，鉴定1个新抗真菌抗肿瘤化合物。进一步采用16S rRNA高变区标签测序技术对一个蚁巢蚂蚁共生微生物进行群落分析，发现放线菌门为第二大共生菌门，包含多种放线菌科。本项目拟对多个蚁巢蚂蚁共生微生物16S rRNA高变区测序分析，明确其放线菌种群组成及丰度，选用专用培养基或特殊筛选方法进行分离、鉴定，并结合快速、高效的新化合物筛选法（LC/MS-PCA）分离新化合物。将纯化、表征为新结构的化合物进行活性测试，以期获得具有新结构特征或新作用特点活性化合物。

昆虫共生放线菌作为一类特境微生物，合成多种新奇结构化合物，为发现新的微生物天然产物药物提供可能。本研究以拟黑多刺蚁为研究对象，通过高通量测序明确其共生微生物的菌群结构，发现可产生丰富代谢产物的放线菌门为其第二大共生菌门，包含类诺卡氏菌菌科，诺卡式菌科及链霉菌科等多种放线菌科。进而采用专用的培养基和筛选方法对其共生放线菌科进行分离和鉴定，共分离不同放线菌327株，鉴定了放线菌新种13株。对分离的菌株进行活性检测，发现活性菌67株。对部分新种及活性菌进行化合物分离，共分离活性化合物28个，其中新化合物5个。以上研究说明昆虫可作为寻找不寻常微生物的重要资源库，为后续研究和开发昆虫共生微生物资源提供了基础和保证。本研究共发表SCI文章13篇，授权发明专利1项。