DEMED · Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2020-12-01 → 2023-05-19
- EU contribution
- €162,806
- Participants
- 2
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays
Issue being addressed and importance: Directed evolution of metalloenzymes based on directed eletron transfer of enzymes and interfaces are missing in literature, in particularly, to direct metalloenzymes to fit the best possible their electron transfer environment. However, it is important for metalloenzyme applications, for example in enzymatic biofuel cells. Overall objectives: 1. Metalloenzyme expression in a way suitable for high throughput screening. 2. Electrochemical droplet microarray design and manufacture. 3. Directed evolution of metalloenzymes on electrochemical droplet microarray. The action was planted with success. However, more time is needed to achieve the final goal that a metalloenzyme perform its best possible in application environment. Time is also needed to patent and publish the screening platform, and the works could be done on the new platform as well.
Data: CORDIS, © European Union
Project objective
The goal of this Marie Curie Individual fellowship proposal is to establish directed evolution of redox enzymes by means of electrochemical microarrays (DEMED) to enable the direct screening of the enzyme properties desired for their application in electrochemical devices. An O2 reducing metalloenzyme for implementation in biocathodes of H2/O2 enzymatic fuel cells will serve as model system to demonstrate that directed evolution of such redox enzymes screened by electrochemical droplet microarray is advantageous to specifically improve biofuel cell performances. The selected metalloenzyme is rubredoxin: oxygen oxidoreductase (ROO), which has never been applied to H2/O2 enzymatic fuel cells so far. First, ROO gene will be cloned and its random mutagenesis library will be synthesized. Second, the electrochemical droplet microarray will be adapted to enable the screening of the desired properties of the metalloenzyme. Third, electrochemical directed evolution of ROO will be carried out. Finally, the interface of ROO and electrode based on redox active polymers will be co-evolved with ROO to achieve high electron transfer rates to the enzyme and thus enable the fabrication of a high performance biocathode. It is expected that this project will have a groundbreaking on directed evolution of metalloenzymes for their practical implementation in electrochemical devices.
Original text from CORDIS.
Participants
Links
Data: CORDIS, © European Union