项目名称: 基于土壤微生物宏基因组发掘新颖活性卤化天然产物
项目编号: No.31500035
项目类型: 青年科学基金项目
立项/批准年度: 2016
项目学科: 生物科学
项目作者: 曹明明
作者单位: 南京农业大学
项目金额: 20万元
中文摘要: 土壤微生物是抗菌活性分子的重要来源。然而只有1%环境微生物才能在常规实验条件下得到成功培养,这致使传统的通过纯化菌株和扩大发酵来发现活性分子的方法受到限制。获取土壤微生物总DNA、克隆天然产物的生合成基因簇、并使其在可培养宿主中表达的宏基因组学技术,为发掘新颖抗菌活性分子提供了一种新方法。 卤化天然产物被认为是经卤化酶参与合成的,许多天然来源的抗生素分子即为卤化物,卤原子的引入常常会提高相应生物活性。前期用宏基因组手段从土壤微生物获得了一批包含卤化酶基因的大型DNA片段,并转入了异源宿主,其表达产物表现出了抗菌活性。 本课题拟在前期工作基础之上,运用宏基因组技术筛选卤化酶基因,挖掘相应卤化物的生物合成基因簇;接合现代化分离纯化技术和结构鉴定手段,获得具有抗菌活性的卤化物。
中文关键词: 次级代谢产物;抗生素;宏基因组;合成基因簇;卤化酶
英文摘要: Soil-microorganisms is an important resource for antibiotics. However, only 1% of environment microbes can be cultured in normal conditions, which limited the discovery of bioactive natural products by traditional cultural-depended method. Metagenomic technology provided a cultural-independed method to access the collective genomes of environmental bacterial populations, and to explore the small molecule biosynthetic gene clusters. Biosynthesis gene clusters of halogenated natural products involved halogenase, and halometabolites favored antibiotics. Large DNA fragments harboring halogenase sequences were obtained in our previous work from soil-microorganisms by metagenomic technology, and their heterologous expression products showed anti-bacterial activities. On the basis of previous work, this project aimed to: screen halogenase genes and biosynthesis gene clusters, purify and elucidate target structures, and obtain anti-bacterial active halometabolites.
英文关键词: secondary metabolites;antibiotics;metagenome;biosynthesis gene cluster;halogenase gene