High-yield recombinant protein expression system for Pseudomonas

Keywords: recombinant protein expression, Pseudomonas, synthetic biology, microbial fermentation.

Pseudomonas putida has emerged as a valuable host for recombinant protein production in industrial biotechnology, thanks to its highly versatile metabolism that enables among others cultivation using waste streams or the production of toxic compounds. However, achieving high yields of recombinant proteins depends heavily on the efficient overexpression of the target gene. In Escherichia coli, this is typically accomplished using the phage T7-based pET expression system. While the T7 RNA polymerase (RNAP) is a powerful tool for synthetic biology and protein production across various microbial hosts, its application in Pseudomonas is severely limited due to cytotoxic effects.

We have developed pPUT, an optimized expression system tailored for Pseudomonas species. This system enables robust overexpression of recombinant proteins, achieving a dynamic expression range of up to 200-fold. Additionally, pPUT incorporates a host RNAP inhibitor that effectively decouples cell growth from product formation, resulting in a further 20% increase in protein yield.

Applications & market opportunities

Our technology has broad applicability in microbial fermentations and beyond. The pPUT system enables the development of metabolically versatile and robust bacterial production hosts, offering a compelling alternative to E. coli without requiring a shift to (more costly and complex) eukaryotic model systems.

Collaboration sought

This technology was developed at KU Leuven in the Laboratory of Gene Technology under the direction of Professor Rob Lavigne. We are looking for industrial partners to collaborate with and/or to license-in the technology.