Objective
The proposed research project aims at implementing enzymatic activity in a de novo designed, unbiased protein scaffold. First, simplified active site arrangements deduced from two previously evolved model enzymes (Kemp eliminase and retro-aldolase) will be implanted in the scaffold. This will allow evaluating the extent to which a fully computationally designed and naïve protein can be functionalized and evolved. A recently established fluorescence-based microfluidics setup will be utilized to screen large DNA libraries of several million clones per round of laboratory evolution. The artificial protein scaffold, kindly provided by Prof. David Baker (University of Washington), has been designed to adopt a minimalist TIM barrel fold, which is the most abundant and diversely evolved protein fold found for natural enzymes. Here, the substrate binding pocket is usually formed by an extended loop region on one side of the scaffold, which is not yet present in the naïve, designed variant. Thus, in the second step, I will generate a randomized library of loop fragments, insert into the scaffold and screen for improved activity. This approach can be extended from the retro-aldolase model reaction towards synthetic aldolases that catalyze the stereospecific formation of a new carbon-carbon bond. Ultimately, the aim is to evaluate whether loop libraries are an appropriate tool to broaden the substrate scope of these enzymes. Furthermore, the proposal contains a second, independent approach to equip the artificial scaffold with novel enzymatic functionality. Here, I plan to design a covalent dimer of two artificial TIM barrels carrying a cofactor-dependent active site in the dimeric interface.
This work will not only generate fundamental insight into the evolution of catalytic activity, it also has great potential to contribute to the development of general strategies for creating enzymes with novel functionality, and thus, prospective applications in industry or medicine.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences physical sciences classical mechanics fluid mechanics microfluidics
- engineering and technology materials engineering crystals
- natural sciences biological sciences genetics DNA
- natural sciences chemical sciences catalysis biocatalysis
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
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H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions
MAIN PROGRAMME
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H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility
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Topic(s)
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Funding Scheme
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
MSCA-IF-EF-ST - Standard EF
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Call for proposal
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
(opens in new window) H2020-MSCA-IF-2015
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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.
8092 Zuerich
Switzerland
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.