Academic Research

The biobased production of recombinant proteins has great potential for future applications and circularized economies. In recent years, synthetic biology toolboxes have been developed for the use of Gram-negative bacteria as production chassis. However, a cell lysis step is often required due to the protein production taking place intracellularly. While a variety of conventional methods exist, many cause impurities and could damage the protein product. This makes production expensive and time-consuming.

The flagellar Type III Secretion System (fT3SS) is a bacterial secretion system found in both Gram-positive and Gram-negative bacteria. This naturally secretes flagellin at a high rate, which is then assembled outside of the cell to form the flagella. Unlike the injectisome, the fT3SS is non-pathogenic. Together with its ability to secrete recombinant proteins, this makes it an interesting secretion platform for the industry.

While much is known about the fT3SS, it is tightly regulated and poorly characterized with regard to the export of recombinant proteins. Here, we aim to develop it into an efficient secretion platform. This is done by characterizing and optimizing secretion drivers, as well as engineering the fT3SS regulation. Combined with engineering strains that can synthesize recombinant proteins in vivo, this creates a high-yield, low-cost bacterial production platform. Such platforms can lower barriers for the production of specific proteins, as well as accelerate the development of a quick, affordable pipeline for new industrial and therapeutic solutions.

The internship will build upon preliminary work pertaining to the general project. The intern will be trained in the full experimental pipeline from literature review to validation. This includes designing, constructing, and cloning new genetic constructs, as well as performing assays to characterize the system.
Techniques mastered by the end of the internship include, but are not limited to: Gibson assembly, one-step mutagenesis, bacterial transformation, and protein detection assays. Exact tasks will depend on the state of the project at the time of arrival.

Master's students