四氢呋喃（THF）是一种大宗工业环醚类污染物。假诺卡氏菌CB1190能以THF为唯一碳源能源进行生长。基因簇thmADBC被推测编码一种新的多组分含双核铁单加氧酶，全细胞生物转化证明其能转化THF，但产物未鉴定，更无酶学性质研究的报道。该菌株THF代谢中的中间产物及除thmADBC外的代谢基因也从未被鉴定。本研究将首次纯化ThmADBC，鉴定THF催化产物，研究其理化特征、底物范围及酶学动力学参数等性质；同时，基于该菌株已有的基因组和转录组数据，本研究将通过生物信息学分析和文库构建来获得完整的THF降解基因；基因敲低、代谢产物积累分析结合基因体外表达、酶活测定等方法将用于分析这些基因的功能，从而在分子、生化和遗传学水平上阐明THF完整的代谢途径。该研究将有助于理解微生物代谢有机污染物的多样性，推动此多组分加氧酶家族的理论研究，同时为THF的生物修复提供理论基础和技术指导。

The cyclic ether pollutant tetrahydrofuran (THF) is used in large amounts for various industrial purposes. Pseudonocardia dioxanivorans strain CB1190 has been identified to utilize THF as a sole source of carbon and energy. It was predicted that the four-gene cluster thmADBC of strain CB1190 is involved in assembly of a new member of multi-component diiron monooxygenases. Transformation of THF was observed by whole cells of Rhodococcus jostii RHA1 harboring thmADBC, but the oxidation product has not yet been identified and no biochemical properties of this enzyme have been characterized. In addition, no metabolic intermediate or catabolic gene (except for thmADBC) involved in THF biodegradation by strain CB1190 has ever been identified. This study will present the first purification of ThmADBC. Based on this, the product of oxidized THF will be identified and the characteristics of the multi-component enzyme, such as the biochemical and physical properties of each component, substrate range and kinetic parameters, will be examined. Besides that, based on the reported genomic and transcriptomic data of strain CB1190, the complete THF catabolic genes will be obtained by bioinformatics approaches and genomic library construction. The function of each gene will be identified through methods of gene knockdown, metabolic intermediates identification, enzyme expression in vitro, and enzyme assay. And in this study, the pathway and mechanisms of THF metabolism in strain CB1190 will be elucidated at molecular, biochemical, and genetic levels. The data present here will be greatly helpful to understand the diverse capabilities for the microbial degradation mechanisms of organic pollutants and the functional characterization of thmADBC genes will certainly promote the theoretical study of this family of multi-component oxygenases. Meanwhile, it will provide a theoretical basis and technical instruction for bioremediation of tetrahydrofuran in the contaminated environment.