恶臭假单胞菌Pseudomonas putida F1石油烃降解细胞工厂的构建与应用

目前有限的微生物资源和基因元件数量限制了石油烃的降解及其生物修复工作的开展。以现有降解菌株作为底盘细胞，插入不同功能基因元件，构建全新具备自杀控制功能的石油烃定向降解细胞工厂，对石油污染的生物修复具有重要意义。本项目拟利用合成生物学、生物信息学、分子生物学等手段开展石油烃降解细胞工厂的构建工作，突破目前石油烃降解功能菌自然进化难以满足环境需求的局限性。1）以恶臭假单胞菌Pseudomonas putida F1为底盘细胞，人工设计、合成和组装石油烃降解相关基因元件以及自杀基因元件，构建新型细胞工厂；2）探究不同基因元件对石油烃降解的耦合作用机理及其与底盘细胞的适配机制，筛选安全高效的基因工程菌株；3）利用转录组学、蛋白质组学等多组学分析，明晰其在石油污染环境中的生存、繁殖、降解等行为特征，评价外源重组基因的转移能力和环境安全性及健康风险。这一研究将为石油污染的生物修复提供重要的参考价值。

At present, the limited microbial resources and genetic elements restrict the development of petroleum hydrocarbon degradation and bioremediation. Using existing degrading strains as platform cells, inserting different functional gene elements to construct a brand-new petroleum hydrocarbon directed degrading cell factory with suicide control function is of great significance to the bioremediation of petroleum pollution. This project intends to combine synthetic biology, bioinformatics, molecular biology and other methods to carry out the construction of petroleum hydrocarbon degradation cell factories, breaking through the limitations of the current natural evolution of petroleum hydrocarbon degradation functional bacteria that is difficult to meet environmental needs. 1) To use Pseudomonas putida F1 as the platform cell, artificially design, synthesize and assemble petroleum hydrocarbon degradation-related genetic elements and suicide genetic elements to construct a new type of cell factory; 2) To explore the coupling effect of different genetic elements on petroleum hydrocarbon degradation mechanism and its adaptation mechanism to platform cells to screen safe and efficient genetically engineered strains; 3) To apply transcriptomics and proteomics analysis to clarify its characteristics including survival, reproduction, degradation and other behaviors in oil-polluted environment, evaluate the transfer capacity of exogenous recombination genes and environmental safety and health risks. This research will provide important reference value for the bioremediation of oil pollution.