Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays

Informazioni relative al progetto

DEMED

ID dell’accordo di sovvenzione: 899066

DOI

10.3030/899066

Progetto chiuso

Data della firma CE 17 Marzo 2020

Data di avvio 1 Dicembre 2020

Data di completamento 19 Maggio 2023

Finanziato da

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Costo totale

€ 162 806,40

Contributo UE

€ 162 806,40

Coordinato da

TECHNISCHE UNIVERSITAET MUENCHEN
Germany

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

Periodo di rendicontazione: 2020-12-01 al 2022-11-30

Sintesi del contesto e degli obiettivi generali del progetto

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.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

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.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

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