About the Project
Recombinant protein production is a major technique used in biotechnology where a host organism is used to synthesise proteins such as drugs, enzymes, or for structural studies. The host we use is the bacterium E. coli, which is good at making relatively simple proteins such as insulin or antibody fragments. Although E. coli cannot perform as many post-translational modifications as mammalian cells, it can form disulphide bonds, which are important for the folding and function of many products.
To form disulphide bonds, the recombinant protein is usually directed to the periplasm (the space between the inner and outer membranes) where the enzymes that create disulphide bonds are located. This means that the protein must pass through the inner membrane, a step which makes processes more complicated and can cause issues with protein misfolding and cell death. We would like to improve this process.
In this project we will explore how we can best improve translocation of recombinant proteins into the periplasm. We have previously developed optimised signal peptides, fermentation processes to improve periplasmic production, and feedback systems to allow us to sense when bacteria are stressed and allow production to be automatically modulated. In this project we will explore these approaches to improve the quantity and quality of accumulated periplasmic protein and de-risk processes to ensure success.
We will use a combination of approaches, grounded in single-cell analysis using flow cytometry, fermentation development, and molecular biology.
This interdisciplinary project will be supervised by Professor Tim Overton, an expert on bioprocess development, single-cell analysis of bacteria, and bacterial physiology, and Professor Owen Thomas, an expert in downstream processing of proteins.
Funding notes:
This is a PhD studentship with the Midlands Integrated Biosciences Training Partnership, funded by BBSRC and in partnership with the University of Warwick, University of Birmingham, Aston University, Harper Adams University, Coventry University, and the University of Leicester.
For more details please visit: https://warwick.ac.uk/fac/cross_fac/mibtp/ or https://www.birmingham.ac.uk/about/college-of-life-and-environmental-sciences/midlands-integrative-biosciences-training-partnership
