Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays

Informations projet

DEMED

N° de convention de subvention: 899066

DOI

10.3030/899066

Projet clôturé

Date de signature de la CE 17 Mars 2020

Date de début 1 Decembre 2020

Date de fin 19 Mai 2023

Financé au titre de

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coût total

€ 162 806,40

Contribution de l’UE

€ 162 806,40

Coordonné par

TECHNISCHE UNIVERSITAET MUENCHEN
Germany

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

Période du rapport: 2020-12-01 au 2022-11-30

Résumé du contexte et des objectifs généraux du projet

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.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

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.

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

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