Summary
In recent years, membraneless organelles were shown to be intracellular liquid like condensates formed through phase separation. Following their discovery, they have been found across all kingdoms of life. For some of these organelles, a crucial role in cellular physiology has been determined, while for many others their true function remains unresolved. Recently, it has also emerged that the pyrenoid, which concentrates the CO2 fixation machinery of photosynthesis, behaves like a liquid non-membrane bound organelle in green algae. However, the pyrenoid and its forming protein EPYC1 undergo a complicated assembly in vivo, which poses a challenge for the functional understanding, engineering and transplantation of pyrenoids. Here I propose to use a bottom-up strategy to assemble and evolve pyrenoid formation de novo. To paraphrase the renowned physicist Richard Feynman – “What I cannot create, I do not understand” – I plan to mimic the formation of pyrenoids by fusing liquid-liquid phase separating (LLPS) intrinsically disordered peptides (IDP) to RuBisCO to create artificial pyrenoid-like condensates in vitro (objective 1). From the RuBisCO-IDP library, I will select the best performing condensate for transplanting into S. elongatus to generate an artificial pyrenoid. Subsequently, I will compare this strain with the wild-type and a carboxysome knock-out strain (objective 2). Finally, I will evolve the RuBisCO-IDP and the corresponding strain for enhanced photosynthesis (objective 3). My experiments will inform on the evolutionary differences between different carbon concentrating mechanisms (CCM), on the biochemical mechanisms underlying a liquid-liquid phase separated CCM and will show whether and how CCMs can be engineered towards improved photosynthesis. This project will lay the basis to realize strategies for increased CO2 uptake in phototrophs, thereby providing new options for a carbon-neutral bioeconomy and improved food productivity in the future.
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More information & hyperlinks
| Web resources: | https://cordis.europa.eu/project/id/101106795 |
| Start date: | 01-11-2023 |
| End date: | 30-04-2025 |
| Total budget - Public funding: | - 130 385,00 Euro |
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Original description
In recent years, membraneless organelles were shown to be intracellular liquid like condensates formed through phase separation. Following their discovery, they have been found across all kingdoms of life. For some of these organelles, a crucial role in cellular physiology has been determined, while for many others their true function remains unresolved. Recently, it has also emerged that the pyrenoid, which concentrates the CO2 fixation machinery of photosynthesis, behaves like a liquid non-membrane bound organelle in green algae. However, the pyrenoid and its forming protein EPYC1 undergo a complicated assembly in vivo, which poses a challenge for the functional understanding, engineering and transplantation of pyrenoids. Here I propose to use a bottom-up strategy to assemble and evolve pyrenoid formation de novo. To paraphrase the renowned physicist Richard Feynman – “What I cannot create, I do not understand” – I plan to mimic the formation of pyrenoids by fusing liquid-liquid phase separating (LLPS) intrinsically disordered peptides (IDP) to RuBisCO to create artificial pyrenoid-like condensates in vitro (objective 1). From the RuBisCO-IDP library, I will select the best performing condensate for transplanting into S. elongatus to generate an artificial pyrenoid. Subsequently, I will compare this strain with the wild-type and a carboxysome knock-out strain (objective 2). Finally, I will evolve the RuBisCO-IDP and the corresponding strain for enhanced photosynthesis (objective 3). My experiments will inform on the evolutionary differences between different carbon concentrating mechanisms (CCM), on the biochemical mechanisms underlying a liquid-liquid phase separated CCM and will show whether and how CCMs can be engineered towards improved photosynthesis. This project will lay the basis to realize strategies for increased CO2 uptake in phototrophs, thereby providing new options for a carbon-neutral bioeconomy and improved food productivity in the future.Status
SIGNEDCall topic
HORIZON-MSCA-2022-PF-01-01Update Date
31-07-2023
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