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Continuous Enzyme Evolution – solving bottlenecks in enzyme engineering to design next-generation biocatalysts

Project description

New methods for next-generation biocatalyst design

Directed evolution has advanced enzyme use in industry by enabling customisation for specific needs. Continuous evolution (CE) methods streamline this process within a replicating organism, allowing for the rapid development of efficient enzymes. However, most approaches focus on model enzymes with limited industrial application. The ERC-funded ContiZymes project will expand the potential of CE by establishing a low-tech circular economy platform for biocatalysts. It will integrate a versatile selection system with gene diversification strategies and an autonomous setup for continuous bacterial growth. This integration will enable the exploration of evolutionary paths to address challenges in C-C, C-halogen, and C-N bond-forming reactions, engineering enzymes while mapping their fitness landscapes to facilitate on-demand design of next-generation biocatalysts.

Objective

Directed evolution has revolutionized the application of enzymes in industrial settings by allowing users to tailor the properties and activities of biocatalysts to their needs. But classic directed evolution is notoriously labor- and time-intensive, as it manually stages mutation, selection, and amplification cycles. In contrast, continuous evolution (CE) approaches aim to achieve these steps within a replicating organism, making it possible to engineer efficient enzymes in a matter of days rather than months or years. Unfortunately, current CE approaches are typically applicable only to model enzymes with little industrial value.

To unleash the full potential of CE, we will develop a scalable, low-tech CE platform, which is readily applicable to biocatalysts that provide value-added products. Toward this end, we will merge a versatile selection system we recently developed with strategies to diversify the genes of targeted enzymes in vivo and an autonomous setup to grow bacterial populations continuously. Combined the resulting CE-platform will enable us to engineer biocatalysts along many and long evolutionary trajectories. Moreover, analyzing the fate of these populations by sequencing will allow us to map the sequence-structure-function relationships of these biocatalysts. Based on the systematic datasets generated in these efforts, we will train machine-learning (ML)-models to predict functional sequences. Lastly, in a ML-directed CE approach, we will establish a design-build-test-learn cycle to improve models and guide CEs toward promising, but otherwise inaccessible sequence spaces.

Overall, ContiZymes will overcome unaddressed challenges associated with the application of biocatalysts that promote sought-after C-C, C-halogen, and C-N-bond forming reactions. We will not only engineer these enzymes at an unprecedented rate and scale, but also map their fitness landscapes and take a critical step toward the on-demand design of next-generation biocatalyst

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Programme(s)

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Topic(s)

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HORIZON-ERC - HORIZON ERC Grants

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Call for proposal

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(opens in new window) ERC-2023-COG

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Host institution

RIJKSUNIVERSITEIT GRONINGEN
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 1 999 991,00
Address
Broerstraat 5
9712CP Groningen
Netherlands

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Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 1 999 991,00

Beneficiaries (1)

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