本文選題：深海宏基因組 + 次生代謝產(chǎn)物�。� 參考：《廈門大學(xué)》2014年碩士論文

【摘要】：在微生物代謝產(chǎn)物及生合成途徑的研究中,由于絕大多數(shù)微生物很難或不能通過分離培養(yǎng)方法獲得純培養(yǎng),因而無法正確認(rèn)識環(huán)境中微生物基因及其編碼生合成代謝產(chǎn)物的多樣性,無法全面認(rèn)識自然界物種的多樣性。因此,非可培養(yǎng)方法是獲知絕大多數(shù)微生物的基因多樣性及代謝多樣性的主要方法。通過直接從環(huán)境樣品中提取微生物的總DNA,并克隆到合適的可培養(yǎng)微生物宿主中,利用基因組學(xué)的研究策略研究環(huán)境樣品所包含的全部微生物的遺傳組成,已成為海洋等極端環(huán)境樣品的重要分析方法。 本文首先對源于深海沉積物樣本的宏基因組文庫中具有細胞毒活性的克隆子進行了化學(xué)篩選,通過與宿主大腸桿菌對比,發(fā)現(xiàn)克隆子10-1及25D7能夠產(chǎn)生差異代謝產(chǎn)物。進而大規(guī)模發(fā)酵宿主大腸桿菌及10-1,對其代謝產(chǎn)物進行提取、分離與結(jié)構(gòu)鑒定,得到18個化合物,為6個吲哚生物堿,5個氯霉素衍生物,7個環(huán)二肽。其中10-1產(chǎn)生的3-羥基-2-吲哚酮類化合物和靛玉紅為差異成分,其生合成途徑可能是加氧酶功能基因作用下氧化底物吲哚形成靛紅,進而縮合成靛玉紅。該生合成途徑通過添加底物5-溴吲哚培養(yǎng)10-1得到證實。克隆子25D7的代謝產(chǎn)物的研究直接通過5-溴吲哚底物添加方式進行。發(fā)酵液在LC-MS指導(dǎo)下定向分離得到4個溴代產(chǎn)物。其中兩個新的溴代雙吲哚衍生物生合成途徑是加氧酶基因和羥化酶基因表達產(chǎn)生2,3-吲哚-二酮和7-羥基吲哚,再縮合成雙吲哚衍生物。通過上述研究,共從兩株深海宏基因組克隆子和宿主發(fā)酵物中獲得23個化合物,其中新化合物2個；驗證了4個差異代謝物的生合成途徑,為進一步開發(fā)和利用深海功能基因奠定了基礎(chǔ)。
[Abstract]:In the study of microbial metabolites and biosynthesis pathway, the majority of microbes are difficult or unable to obtain pure culture by the method of isolation and culture. Therefore, it is impossible to correctly understand the diversity of microbial genes and their encoded metabolites in the environment, and to fully understand the diversity of natural species. Therefore, the non-culturable method is the main way to know the gene diversity and metabolic diversity of most microorganisms. By extracting the total DNA of microbes directly from environmental samples and cloning them into suitable culturable microorganism hosts, the genetic composition of all microbes contained in environmental samples was studied using genomics research strategies. It has become an important analytical method for extreme environmental samples such as the ocean. In this paper, the cytotoxic clones from macrogenomic library derived from deep-sea sediment samples were chemically screened. By comparison with host Escherichia coli, it was found that clone 10-1 and 25D7 could produce different metabolites. The metabolites of Escherichia coli and 10-1 were extracted, isolated and identified, and 18 compounds were obtained, including 6 indole alkaloids, 5 chloramphenicol derivatives and 7 cyclic dipeptides. The 3-hydroxy-2-indole compounds produced by 10-1 and indirubin were different components. The pathway of their synthesis may be the oxidation of indole to indirubin under the action of oxygenase function gene, and then the synthesis of indirubin. The biosynthesis pathway was confirmed by adding 5-bromoindole to culture 10-1. The metabolites of clone 25D7 were studied directly by adding 5-bromoindole substrate. Under the guidance of LC-MS, four brominated brominated products were obtained. Two new brominated diindole derivatives were synthesized by the expression of oxygenase gene and hydroxylase gene to produce 2-indole-3-diketone and 7-hydroxyindole, and then condensed to bis (indole) derivatives. Based on the above studies, 23 compounds were obtained from two deep-sea macrogenomic clones and host fermentations, 2 of which were new compounds, which verified the biosynthesis pathway of 4 differentially metabolites. It lays a foundation for the further development and utilization of deep-sea functional genes.
【學(xué)位授予單位】：廈門大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R915

【參考文獻】

相關(guān)期刊論文 前3條

1 孫宇輝,周秀芬,涂國全,鄧子新;南昌鏈霉菌代謝產(chǎn)物與代謝工程研究現(xiàn)狀的回顧與展望[J];生命科學(xué);2005年03期

2 左佃光;馬俊英;王博;黃洪波;劉靜;張云;鞠建華;;非核糖體肽類化合物的組合生物合成策略研究進展[J];中國抗生素雜志;2012年03期

3 羅坤;趙志祥;蘆曉飛;黎定軍;謝丙炎;;拮抗南方根結(jié)線蟲PKS基因的篩選及活性評價[J];植物保護;2010年05期

，

本文編號：1782435