About the project – University of Copenhagen

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About the Project

Bio-active terpenoids are complex plant compounds with a wide range of valuable properties – for instance as anti-cancer agents, and potentially as hormonal regulators or antibacterial agents against multidrug-resistant bacteria.

Unfortunately, terpenoids are extremely difficult or impossible to synthesise by chemical means, and only produced in plants in minute amounts. Our goal for this project is to produce high amounts of valuable terpenoids. Therefore we will transfer enzymatic pathways for specific terpenoids from higher plants to cyanobacteria and moss, and these pathways will be optimised by re-targeting them to the chloroplasts and directly coupling them to the photosynthetic system – leading to light-driven production.

The key enzymes in the synthesis of terpenoids are chloroplastic terpenoid synthases and a series of cytochrome P450s (in short: P450s) that are located in the endoplasmic reticulum. Using the “share your parts” principle of synthetic biology we will engineer photosynthetic organisms to produce large quantities of specific terpenoids in photobioreactors. The new approaches use recent technology in which P450s were relocated to the chloroplast and directly driven by photosynthetic electron transport for the first time. The project will develop a suite of compound-specific production platforms using selected combinations of terpenoid synthases and P450s.

Chloroplast bioengineering. A. Structure of the diterpenoid ingenol-3-angelate. B. Photosynthesis and biosynthesis of specialised bioactive compounds are separated in the chloroplast and endoplasmic reticulum, respectively. C. Using bioengineering we aim at co-localizing the biosynthetic pathway of ingenol-3-angelate together with the photosynthetic complexes in the chloroplasts of moss and in cyanobacteria. Note that the same principles can be used to target P450s into the cyanobacterial thylakoid membrane and likewise drive the reactions using solar energy.

Production systems will be developed and optimised in chloroplasts, and then transferred to cyanobacteria and moss strains for growth in closed photobioreactors. The program will involve extensive collaboration with groups from the Univ. of California, Berkeley, and Univ. of Kent, who provide world-leading expertise in synthetic biology and protein targeting, and a series of industrial collaborators will help to develop and validate the photobioreactor production platforms under full production conditions. The overall result will be the development of novel, carbon-neutral production platforms for complex terpenoids, the creation of a generic system that can be adapted for future terpenoids, and the establishment of state-of-the-art photobioreactor technology in Denmark.