Project properties

Title Establishing anaerobic activity of heterologous PFOR in E. coli
Group Systems and Synthetic Biology
Project type thesis
Credits 36
Supervisor(s) Jenny Bakker, Maria Suarez Diez
Examiner(s) Prof. Maria Suarez Diez (SSB) Dr. Rob Smith
Contact info Robert1.smith@wur.nl
Begin date 2025/06/05
End date
Description In this project, you will work on establishing the activity of heterologous pyruvate::ferredoxin oxidoreductases (PFOR). These enzymes are found in many different organisms, and our host organism, E. coli, natively also has one. However, the one from E. coli takes flavodoxin instead of ferredoxin. We aim to increase the energy conservation in E. coli, and to achieve this, the ferredoxin produced by PFOR should be oxidised by Rnf, an enzyme found in acetogens. Rnf will produce a proton motive force when it oxidises ferredoxin. It is crucial that the correct type of ferredoxin is produced so that it can interact with the Rnf. Therefore, heterologous PFOR activity in E. coli needs to be established. Growth-coupled biosensors have been developed and characterised previously to test the heterologous PFOR activity.
Used skills Understanding of metabolic engineering strategies for microorganisms.
Requirements Basic microbiology laboratory skills