Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays

Project Information

DEMED

Grant agreement ID: 899066

DOI

10.3030/899066

Project closed

EC signature date 17 March 2020

Start date 1 December 2020

End date 19 May 2023

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 162 806,40

EU contribution

€ 162 806,40

Coordinated by

TECHNISCHE UNIVERSITAET MUENCHEN
Germany

Periodic Reporting for period 1 - DEMED (Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays)

Reporting period: 2020-12-01 to 2022-11-30

Summary of the context and overall objectives of the project

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.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

Metalloenzymes were expressed successfully. Surface display systems of microorganisms were employed in the protein expression systems to fit high throughput screening. Electrochemical droplet microarrays were designed and made for enzyme mutant screening. Analytical standards of microarrays were defined. The microarray was integrated into a newly-built high throughput screening platform. Preliminary screening of metalloenzymes on electrochemical microarrays were carried out.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

New collaborations were established to complete the project. Enzyme activities will be inspected further by mass spectroscopies. Cell sorting and single-cell sequencing will be used to analyze droplet mutants at genomic level. This project is filling up a scientific gap. Herein, the achievements of the project will have a general impact on academic and industrial world.

Directed evolution on electrochemical droplet microarray