Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays

Projektinformationen

DEMED

ID Finanzhilfevereinbarung: 899066

DOI

10.3030/899066

Projekt abgeschlossen

EK-Unterschriftsdatum 17 März 2020

Startdatum 1 Dezember 2020

Enddatum 19 Mai 2023

Finanziert unter

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Gesamtkosten

€ 162 806,40

EU-Beitrag

€ 162 806,40

Koordiniert durch

TECHNISCHE UNIVERSITAET MUENCHEN
Germany

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

Berichtszeitraum: 2020-12-01 bis 2022-11-30

Zusammenfassung vom Kontext und den Gesamtzielen des Projekts

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.

Arbeit, die ab Beginn des Projekts bis zum Ende des durch den Bericht erfassten Berichtszeitraums geleistet wurde, und die wichtigsten bis dahin erzielten Ergebnisse

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.

Fortschritte, die über den aktuellen Stand der Technik hinausgehen und voraussichtliche potenzielle Auswirkungen (einschließlich der bis dato erzielten sozioökonomischen Auswirkungen und weiter gefassten gesellschaftlichen Auswirkungen des Projekts)

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