酞酸酯被广泛用作塑化剂及化工产品的添加剂，在环境中分布广泛，自然降解极慢，严重危害环境安全和人类健康，微生物代谢是酞酸酯降解的主要途径。申请者已分离到酞酸酯高效降解菌Mycobacterium sp.YC-RL4、Gordonia alkanivorans YC-RL2，并完成了YC-RL4对酞酸酯的降解特性、酯酶基因克隆及鉴定、全基因组测序等工作。本项目拟在此基础上进行以下研究：（1）进行酞酸酯酶DPH1、DPH2和MPH1酶学特征与酶三维结构分析以解析其催化的分子机制；（2）通过酞酸酯诱导下转录组和蛋白组的研究鉴定酞酸酯代谢相关基因；（3）用RT-qPCR、CRISPR/Cas9基因敲除验证候选基因，BIAcore分析调控蛋白与DNA的相互作用，最终构建酞酸酯降解的完整遗传调控网络，全面深入揭示酞酸酯的降解机理，研究结果将对酞酸酯降解资源的开发与可持续应用具有重要的理论和实践意义。

Phthalate is widely applied as plasticizer and additive of other chemical products. Phthalate is universally distributed in the environment, and is extremely difficult to degrade under natural conditions, which has done damage to environment safety and health of human. Microbial metabolism is the main approach for elimination of phthalate. Some strains including Mycobacterium sp. YC-RL4 and Gordonia alkanivorans YC-RL2 isolated previously by the applicant were capable of degrading phthalate efficiently. Based on the accomplished degradation properties, cloning and identification of esterase genes and complete genome sequencing of strain YC-RL4, the following experiments will be performed： (1) demonstrating the efficient catalytic mechanism of the esterases DPH1, DPH2 and MPH1 by analyzing the enzymatic characteristics and 3D structures; (2)screening genes involved in phthalate metabolism by analysis of the transcriptome and proteome of strain YC-RL4 under phthalate induction; (3)verifying the candidate genes by RT-qPCR and gene knockout using CRISPR/Cas9, and analyzing the interaction of regulatory proteins and DNA. In conclusion, the model of phthalate metabolism and regulation network will be built, and the molecular mechanism of phthalate degradation will be proposed. The results will be theoretically and practically significant for the application and sustainable development of resources of phthalate degradation.