Squalene, a universal precursor of various bioactive compounds, has unique physical and chemical properties, making it beneficial to human. Sharks' liver oil is the richest natural origin of squalene. However, this origin is limited because of epidemic marine pollution, overfishing of sharks and the international concern on shark protection. The goal of this research is to produce squalene using E.coli, the most common microbial factory. To achieve this, we need to introduce exogenous squalene synthase (SQS) into E.coli. Firstly, we compared the yield using SQSs from different species. To facilitate this, CrtN, turning colorless squalene into yellow ß-carotene, is introduced. Secondly, an exogenous MVA pathway is introduced and a more effective MEP pathway is developed via overexpression of key regulatory enzymes. Both pathways produce the precursor of squalene in E.coli and eventually improve the squalene yield. This research provides an alternative to squalene production, bringing positive effects on shark protection.
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