Knowledge Resource Center for Ecological Environment in Arid Area
| 南中国海和新疆沙漠菌种多样性和一株链霉菌次级代谢产物研究 | |
| 其他题名 | THE STUDY OF BACTERIA DIVERSITY FROM THE SOUTH CHINA SEA AND XINJIANG DESERT AND CHEMICAL INVESTIGATION OF ONE STREPTOMYCES STRAIN |
| 杨娜 | |
| 出版年 | 2014 |
| 学位类型 | 博士 |
| 导师 | 张立新 |
| 学位授予单位 | 中国科学院大学 |
| 中文摘要 | 基于对陆生微生物天然产物的重复发现率越来越高，故而将研究对象转为海洋与沙漠等特殊生境来源的微生物菌株。南中国海与新疆沙漠为典型的特殊生境，蕴含了多样和独特的微生物资源，是研究微生物资源与天然产物的宝库。结核病 (tuberculosis, TB) 和耐甲氧西林金黄色葡萄球菌(MRSA)感染导致的死亡率居高不下，已有药物远不能满足现有需求，对抗感染药物的研发迫在眉睫。本实验室已建立起基于牛结核分枝杆菌(Bacillus Calmette-Guérin, BCG)和MRSA的高通量筛选模型，对天然产物及化合物进行初步的活性筛选。 本论文主要包括从纯培养的角度在多相分类学的指导下对南中国海沉积物和新疆沙漠样品进行菌种分离，经去重复化后，从17个样品中获得放线菌菌株243株，分属于13个菌属(Streptomyces，Nocardiopsis，Prauserella，Tsukamurella，Salinispora，Micromonospora，Verrucosispora，Promicromonospora，Plantactinospora, Saccharopolyspora, Saccharomonospora，Actinocorallia, Nocardia)；1个嗜碱菌属Amphibacillus菌株；2个中度嗜盐菌属Gracilibacillus和Salinibacillus菌株，包括10株潜在新分类单元的菌株。其中Plantactinospora和Amphibacillus为首次于海洋生境中发现，Amphibacillus为首次报道具有嗜碱特性的海洋微生物，中度嗜盐菌属Salinibacillus为新疆艾丁湖生境所特有。通过形态学、细胞化学和分子等多相分类学手段对嗜碱和中度嗜盐两个类群的菌株完成菌株系统发育定位和描述工作。初步的研究结果显示，南中国海沉积物和新疆沙漠环境菌株资源具有丰富的多样性，且获得新菌种几率较高，具有很好的开发潜力。 本论文还对获得的放线菌菌株进行了初步的抗草分枝杆菌 (Mycobacterium phlei) 活性筛选，共获得18株活性菌株。除经典的产活性产物较多的Streptomyces菌株外，Micromonospora, Verrucosispora，Salinispora和Saccharopolyspora菌株也被发现具有抗M. phlei活性。结合HPLC和LC-MS对活性菌株进行产物初步分析发现了产rifamycin的Salinispora sp. NHF45，产spinosad的Saccharopolyspora sp. NHF132及其他活性产物的菌株。 本论文还对筛选后得到的一株海洋来源的链霉菌菌株Streptomyces sp. NHF165的次级代谢产物进行了研究和生物活性评价，从中分离纯化得到2个化合物，利用多种波谱技术进行结构鉴定。化合物1为actinonin，首次发现其具有抗MRSA活性(MIC 1.56μg/ml)，化合物2为新结构化合物，验证了开发特殊生境菌株是获得新一代抗感染药物前体的重要基础。 |
| 英文摘要 | Rediscovery rate of bioactive compounds from terrestrial microbes is increasing. Hence microbes from marine and desert habits aroused people’s attention. The South China Sea and Xinjiang desert environments are great sources of novel microorganisms and new active compounds due to their particular ecologies. Tuberculosis (TB) and methicillin-resistant Staphylococcus aureus (MRSA) infection are major fatal infectious diseases in the world. New antibiotics are urgently needed to thwart these pathogens. Our lab has constructed high throughput screening assays based on Bacillus Calmette-Guérin (BCG) and MRSA for investigating novel active compounds. This thesis focused on the study of diversity of microbes derived from South China Sea and Xinjiang desert using culture dependent method guided by microbial taxonomy. Totally 243 actinobacteria were isolated after dereplication and they belonged to 13 different genera (Streptomyces，Nocardiopsis，Prauserella，Tsukamurella，Salinispora，Micromonospora, Verrucosispora, Promicromonospora, Plantactinospora, Saccharopolyspora, Saccharomonospora, Actinocorallia, Nocardia). A group of alkaliphilic bacteria (Amphibacillus) and a group of moderate halophilic bacteria (Gracilibacillus and Salinibacillus) were also isolated from South China Sea and Xinjiang desert respectively. Ten of these isolates were considered novel taxa by current research. It was first successfully got Plantactinospora and Amphibacillus strains from marine environment. Salinibacillus was obligately Aiding lake bacteria genus. Polyphasic taxonomy including morphology, physiological and biochemical tests, chemotaxonomy and molecular characteristics were carried out for alkaliphilic taxa and moderate halophilic taxa. The preliminary investigation results indicated South China Sea and Xinjiang desert are extremely rich reservoir for the isolation of a significant diversity of microbes, including novel species. All isolates were tested for bioactive screening. 18 of these strains showed positive results (Streptomyces，Micromonospora, Verrucosispora，Salinispora and Saccharopolyspora). Though HPLC and LC-MS analysis of the crude extract of bioactive strains, we found strain Salinispora sp. NHF45 producing rifamycin, strain Saccharopolyspora sp. NHF132 producing spinosad and other active compounds. Chemical and biological investigations were conducted on a marine derived strain Streptomyces sp. NHF165. Form the extracts of NHF165, compounds 1 and 2 were isolated. The structures of compounds were elucidated by means of spectroscopic methods. Compound 1 was actinonin and was first reported anti-MRSA activity (MIC 1.56μg/ml) and compound 2 had a novel structure. All these results showed microbes derived from extreme environment are potential sources for active and novel natural products discovery. |
| 中文关键词 | 南中国海 ; 新疆沙漠 ; 放线菌 ; 多相分类学 ; 活性天然产物 |
| 英文关键词 | South China Sea Xinjiang desert actinomycetes polyphasic taxonomy bioactive natural compounds |
| 语种 | 中文 |
| 国家 | 中国 |
| 来源学科分类 | 生物化学与分子生物学 |
| 来源机构 | 中国科学院微生物研究所 |
| 资源类型 | 学位论文 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/287421 |
| 推荐引用方式 GB/T 7714 | 杨娜. 南中国海和新疆沙漠菌种多样性和一株链霉菌次级代谢产物研究[D]. 中国科学院大学,2014. |
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