Engineered Microbial\nConsortium for <i>De Novo</i> Production of Sclareolide

Sclareolide, a natural product with bioactive and fragrant\nproperties,\nis not only utilized in the food, healthcare, and cosmetics industries\nbut also serves as a precursor for the production of ambroxide and\nsome bioactive compounds. Currently, there are three primary methods\nfor producing sclareolide: direct extraction from plants, chemical\nsynthesis using sclareol as a precursor, and the biotransformation\nof sclareol. Here, we established a platform for producing sclareolide\nthrough a modular coculture system with Saccharomyces\ncerevisiae and Cryptococcus albidus ATCC 20918. S. cerevisiae was engineered\nfor <i>de novo</i> sclareol biosynthesis from glucose, while C. albidus enabled the production of sclareolide\nvia sclareol biotransformation. To enhance the supply of sclareol,\na recombinant yeast strain was constructed through metabolic engineering\nto produce 536.2 mg/L of sclareol. Further improvement of the coculture\nsystem for sclareolide production was achieved by incorporating Triton\nX-100 facilitated intermediate permeability, inoculation proportion\nadjustment, and culture temperature optimization. These refinements\nculminated in a sclareolide yield of 626.3 mg/L. This study presents\na novel streamlined and efficient approach for sclareolide preparation,\nshowcasing the potential of the microbial consortium in sustainable\nbioproduction.