海洋微生物及其天然产物在两种肝癌诱因防治中的功能发掘及作用机理研究
发布时间：2019-01-06 17:46
【摘要】：海洋系统是地球上最大的生态系统,其中蕴含丰富的微生物资源,这些微生物由于生长环境的独特性,包含了许多新型的代谢类型,其中多种微生物的天然产物已成功开发成各种药物前体,为新型活性物质的寻找提供了良好的资源,同时也为工业菌的开发提供了丰富的种质资源库。本研究利用海洋微生物及其天然产物的多样性,筛选抑制肝癌诱因之一丙型肝炎病毒的活性物质,同时筛选能够修复另一肝癌诱因重金属污染的微生物,并初步探究其中的作用机理。丙型肝炎病毒是诱发非A、非B型肝癌的主要因素,目前可用的治疗手段依然有限。本研究针对丙型肝炎病毒与宿主细胞因子的相互作用,构建了两个高通量筛选模型,从海洋微生物天然产物中筛选能够抑制丙型肝炎病的药物前体。其中一个高通量筛选模型是基于丙型肝炎病毒核糖体进入位点IRES与人体翻译起始因子e IF3的相互作用,采用荧光偏振的技术手段,构建模型并进行活性物质筛选;另外一个高通量筛选模型是基于丙型肝炎病毒非结构蛋白NS5A和人体蛋白CypA的相互作用,采用荧光共振能量转移的技术手段,构建模型并同样进行活性物质筛选。从筛选模型中筛选到两个抑制IRES和eIF3互作的活性物质Mucl39526和NP39,以及一个促进IRES和eIF3互作的活性物质HP-3,对他们的作用机理进行了初步的探讨。重金属污染也是诱发肝癌的重要因素之一,而微生物修复技术在重金属污染治理中具有良好的应用前景。本研究利用产碳酸钙细菌能够同时共沉淀重金属离子的作用,产碳酸钙细菌又与尿素的分解代谢密切相关,首先从海底沉积物中分离的细菌中筛选能够分解尿素的细菌,再从中进一步筛选产碳酸钙细菌,挑取代表性产碳酸钙细菌进行进一步的机理探索与重金属离子共沉淀作用研究。采用SEM-EDX、TEM-SAED、XRD、FTIR等技术验证代表菌株的碳酸钙沉淀及重金属离子共沉淀作用,采用电感耦合等离子体发射光谱技术测定代表菌株对钙离子或重金属离子的沉淀效率,并测定了代表菌株的全基因组与转录组,依次初步了解菌株沉淀碳酸钙、耐受重金属及共沉淀重金属离子的机理。结果表明代表菌株Brevibacterium lines 258能够高效沉淀钙离子和重金属离子(铜、锌、锰、锶),且在其基因组中携带多种重金属的抗性基因,其中ABC转运子与菌株的生物修复功能密切相关,为进一步的工业应用提供了一株良好的功能菌株。
[Abstract]:The marine system is the largest ecosystem on earth, which contains abundant microbial resources, which, because of the unique growth environment, contain many new types of metabolism. The natural products of many microbes have been successfully developed into various drug precursors, which provides a good resource for the search of new active substances, and also provides a rich germplasm resource bank for the development of industrial bacteria. In this study, we used the diversity of marine microorganisms and their natural products to screen the active substances of hepatitis C virus, one of the inducers of liver cancer, and to screen the microorganisms that could repair the heavy metal pollution caused by another liver cancer. And explore the mechanism of action. Hepatitis C virus is the main factor to induce non-A-, non-B liver cancer, and the available treatment is still limited. In this study, two high-throughput screening models were established for the interaction between hepatitis C virus and host cytokines. The drug precursors that could inhibit hepatitis C disease were screened from natural products of marine microorganisms. One of the high-throughput screening models was based on the interaction between IRES, a ribosomal entry site of hepatitis C virus, and the human translation initiation factor e IF3. The model was constructed by fluorescence polarization technique and the active substances were screened. Another high-throughput screening model is based on the interaction between hepatitis C virus non-structural protein NS5A and human protein CypA. Two active substances, Mucl39526 and NP39, which inhibit the interaction between IRES and eIF3, and an active substance, HP-3, which promoted the interaction between IRES and eIF3, were screened from the screening model. Heavy metal pollution is also one of the important factors to induce liver cancer, and microbial remediation technology has a good prospect in the treatment of heavy metal pollution. In this study, calcium carbonate producing bacteria can co-precipitate heavy metal ions at the same time, and calcium carbonate producing bacteria are closely related to the catabolism of urea. Then the bacteria producing calcium carbonate were screened and the representative calcium carbonate producing bacteria were selected to study the mechanism and the coprecipitation of heavy metal ions. The calcium carbonate precipitation and heavy metal ion coprecipitation of the representative strain were verified by SEM-EDX,TEM-SAED,XRD,FTIR and other techniques. Inductively coupled plasma emission spectrometry (ICP-AES) was used to determine the precipitation efficiency of calcium or heavy metal ions of the representative strain, and the whole genome and transcription group of the representative strain were determined, and the calcium carbonate precipitated by the strain was preliminarily understood. Mechanism of tolerance to heavy metals and coprecipitation of heavy metal ions. The results showed that the representative strain Brevibacterium lines 258 could precipitate Ca ~ (2 +) and heavy metal ions (Cu, Zn, mn, Sr) efficiently and carry many heavy metal resistance genes in its genome. The ABC transporter was closely related to the bioremediation function of the strain. It provides a good functional strain for further industrial application.
【学位授予单位】：中国科学院大学(中国科学院海洋研究所)
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.7
本文编号：2403119
[Abstract]:The marine system is the largest ecosystem on earth, which contains abundant microbial resources, which, because of the unique growth environment, contain many new types of metabolism. The natural products of many microbes have been successfully developed into various drug precursors, which provides a good resource for the search of new active substances, and also provides a rich germplasm resource bank for the development of industrial bacteria. In this study, we used the diversity of marine microorganisms and their natural products to screen the active substances of hepatitis C virus, one of the inducers of liver cancer, and to screen the microorganisms that could repair the heavy metal pollution caused by another liver cancer. And explore the mechanism of action. Hepatitis C virus is the main factor to induce non-A-, non-B liver cancer, and the available treatment is still limited. In this study, two high-throughput screening models were established for the interaction between hepatitis C virus and host cytokines. The drug precursors that could inhibit hepatitis C disease were screened from natural products of marine microorganisms. One of the high-throughput screening models was based on the interaction between IRES, a ribosomal entry site of hepatitis C virus, and the human translation initiation factor e IF3. The model was constructed by fluorescence polarization technique and the active substances were screened. Another high-throughput screening model is based on the interaction between hepatitis C virus non-structural protein NS5A and human protein CypA. Two active substances, Mucl39526 and NP39, which inhibit the interaction between IRES and eIF3, and an active substance, HP-3, which promoted the interaction between IRES and eIF3, were screened from the screening model. Heavy metal pollution is also one of the important factors to induce liver cancer, and microbial remediation technology has a good prospect in the treatment of heavy metal pollution. In this study, calcium carbonate producing bacteria can co-precipitate heavy metal ions at the same time, and calcium carbonate producing bacteria are closely related to the catabolism of urea. Then the bacteria producing calcium carbonate were screened and the representative calcium carbonate producing bacteria were selected to study the mechanism and the coprecipitation of heavy metal ions. The calcium carbonate precipitation and heavy metal ion coprecipitation of the representative strain were verified by SEM-EDX,TEM-SAED,XRD,FTIR and other techniques. Inductively coupled plasma emission spectrometry (ICP-AES) was used to determine the precipitation efficiency of calcium or heavy metal ions of the representative strain, and the whole genome and transcription group of the representative strain were determined, and the calcium carbonate precipitated by the strain was preliminarily understood. Mechanism of tolerance to heavy metals and coprecipitation of heavy metal ions. The results showed that the representative strain Brevibacterium lines 258 could precipitate Ca ~ (2 +) and heavy metal ions (Cu, Zn, mn, Sr) efficiently and carry many heavy metal resistance genes in its genome. The ABC transporter was closely related to the bioremediation function of the strain. It provides a good functional strain for further industrial application.
【学位授予单位】：中国科学院大学(中国科学院海洋研究所)
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.7
【参考文献】
相关期刊论文 前1条
1 陈亚刚;陈雪梅;张玉刚;龙新宪;;微生物抗重金属的生理机制[J];生物技术通报;2009年10期
，本文编号：2403119
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