Project description
Engineered bacterial platforms synthesise glycolate directly from RuBisCo
Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) is Earth's most abundant enzyme, used by bacteria, algae and terrestrial plants to convert atmospheric CO2 into organic compounds. Despite its critical importance, RuBisCo’s carbon fixation is not completely efficient, often generating a glycolate precursor as a side product. Glycolate is a versatile chemical that can serve as a substrate for other higher value-added products. With the support of the Marie Skłodowska-Curie Actions programme, the ROAD project will harness RuBisCo’s inefficiency, modifying and augmenting it to enable a novel microbial biorefinery for glycolate production directly from CO2 using engineered bacterial platforms.
Objective
Establishing a bio-based economy requires the development of novel biorefineries, where bacterial cell factories are employed for producing added-value compounds from cheap, renewable substrates. CO2 is the ideal feedstock, being the most abundant and virtually unlimited carbon-source on Earth. Novel, highly promising biorefineries aim to utilize genetically engineered bacteria to convert renewable energies and atmospheric CO2 into fuels and chemicals. The enzyme Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is the main responsible of CO2 fixation in the biosphere via the Calvin-Benson-Bassham cycle. Despite its wide distribution in the tree of Life, Rubisco is a rather inaccurate enzyme, presenting a tendency to perform an oxygenation side-reaction which results in the formation of 2-phosphoglycolate (2PG). While re-assimilation of 2PG in central metabolism results in net CO2 loss, it can serve as a precursor of glycolate, an attractive, versatile platform chemical. Here, I propose to exploit the sloppiness of Rubisco to implement a novel biorefinery for glycolate production from CO2. Ultimately, I aim to demonstrate feasible microbial synthesis glycolate, where engineered bacterial platforms synthesize this molecule directly from CO2 (as carbon source) and renewable H2 or formate (as energy source). This overarching goal will be pursued through three different, complementary objectives, including: in vivo screening of new, recently described Rubisco isoforms via ad hoc designed 'selection strain'; in vivo directed evolution of the best performing Rubisco isoforms for improving the inherent oxygenation activity; engineering of natural (Cupriavidus necator) and synthetic (Escherichia coli) bacteria autotrophs as cell factories for this process, which will be tested in gas-controlled lab-scale reactors. Eventually, such a novel biorefinery concept will open unprecedented possibilities for the use of CO2 as feedstock for a true biobased economy.
Fields of science
Not validated
Not validated
- engineering and technologyenvironmental engineeringenergy and fuelsrenewable energy
- natural sciencesbiological sciencesmicrobiologybacteriology
- natural sciencesearth and related environmental sciencesatmospheric sciencesmeteorologybiosphera
- social scienceseconomics and businesseconomicsbioeconomy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
Programme(s)
- HORIZON.1.2 - Marie Skłodowska-Curie Actions (MSCA) Main Programme
Funding Scheme
MSCA-PF - MSCA-PFCoordinator
2800 Kongens Lyngby
Denmark