Directed Evolution of Metalloenzymes through Electrochemical Droplet Microarrays

Información del proyecto

DEMED

Identificador del acuerdo de subvención: 899066

DOI

10.3030/899066

Proyecto cerrado

Fecha de la firma de la CE 17 Marzo 2020

Fecha de inicio 1 Diciembre 2020

Fecha de finalización 19 Mayo 2023

Financiado con arreglo a

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coste total

€ 162 806,40

Aportación de la UE

€ 162 806,40

Coordinado por

TECHNISCHE UNIVERSITAET MUENCHEN
Germany

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

Período documentado: 2020-12-01 hasta 2022-11-30

Resumen del contexto y de los objetivos generales del proyecto

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.

Trabajo realizado desde el comienzo del proyecto hasta el final del período abarcado por el informe y los principales resultados hasta la fecha

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.

Avances que van más allá del estado de la técnica e impacto potencial esperado (incluida la repercusión socioeconómica y las implicaciones sociales más amplias del proyecto hasta la fecha)

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