海洋微生物及其天然產(chǎn)物在兩種肝癌誘因防治中的功能發(fā)掘及作用機理研究
發(fā)布時間：2019-01-06 17:46
【摘要】：海洋系統(tǒng)是地球上最大的生態(tài)系統(tǒng),其中蘊含豐富的微生物資源,這些微生物由于生長環(huán)境的獨特性,包含了許多新型的代謝類型,其中多種微生物的天然產(chǎn)物已成功開發(fā)成各種藥物前體,為新型活性物質(zhì)的尋找提供了良好的資源,同時也為工業(yè)菌的開發(fā)提供了豐富的種質(zhì)資源庫。本研究利用海洋微生物及其天然產(chǎn)物的多樣性,篩選抑制肝癌誘因之一丙型肝炎病毒的活性物質(zhì),同時篩選能夠修復另一肝癌誘因重金屬污染的微生物,并初步探究其中的作用機理。丙型肝炎病毒是誘發(fā)非A、非B型肝癌的主要因素,目前可用的治療手段依然有限。本研究針對丙型肝炎病毒與宿主細胞因子的相互作用,構(gòu)建了兩個高通量篩選模型,從海洋微生物天然產(chǎn)物中篩選能夠抑制丙型肝炎病的藥物前體。其中一個高通量篩選模型是基于丙型肝炎病毒核糖體進入位點IRES與人體翻譯起始因子e IF3的相互作用,采用熒光偏振的技術(shù)手段,構(gòu)建模型并進行活性物質(zhì)篩選;另外一個高通量篩選模型是基于丙型肝炎病毒非結(jié)構(gòu)蛋白NS5A和人體蛋白CypA的相互作用,采用熒光共振能量轉(zhuǎn)移的技術(shù)手段,構(gòu)建模型并同樣進行活性物質(zhì)篩選。從篩選模型中篩選到兩個抑制IRES和eIF3互作的活性物質(zhì)Mucl39526和NP39,以及一個促進IRES和eIF3互作的活性物質(zhì)HP-3,對他們的作用機理進行了初步的探討。重金屬污染也是誘發(fā)肝癌的重要因素之一,而微生物修復技術(shù)在重金屬污染治理中具有良好的應用前景。本研究利用產(chǎn)碳酸鈣細菌能夠同時共沉淀重金屬離子的作用,產(chǎn)碳酸鈣細菌又與尿素的分解代謝密切相關(guān),首先從海底沉積物中分離的細菌中篩選能夠分解尿素的細菌,再從中進一步篩選產(chǎn)碳酸鈣細菌,挑取代表性產(chǎn)碳酸鈣細菌進行進一步的機理探索與重金屬離子共沉淀作用研究。采用SEM-EDX、TEM-SAED、XRD、FTIR等技術(shù)驗證代表菌株的碳酸鈣沉淀及重金屬離子共沉淀作用,采用電感耦合等離子體發(fā)射光譜技術(shù)測定代表菌株對鈣離子或重金屬離子的沉淀效率,并測定了代表菌株的全基因組與轉(zhuǎn)錄組,依次初步了解菌株沉淀碳酸鈣、耐受重金屬及共沉淀重金屬離子的機理。結(jié)果表明代表菌株Brevibacterium lines 258能夠高效沉淀鈣離子和重金屬離子(銅、鋅、錳、鍶),且在其基因組中攜帶多種重金屬的抗性基因,其中ABC轉(zhuǎn)運子與菌株的生物修復功能密切相關(guān),為進一步的工業(yè)應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.
【學位授予單位】：中國科學院大學(中國科學院海洋研究所)
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R735.7
本文編號：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.
【學位授予單位】：中國科學院大學(中國科學院海洋研究所)
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R735.7
【參考文獻】
相關(guān)期刊論文 前1條
1 陳亞剛;陳雪梅;張玉剛;龍新憲;;微生物抗重金屬的生理機制[J];生物技術(shù)通報;2009年10期
，本文編號：2403119
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