Marker-free genome engineering in Cupriavidus necator for sustainable protein production
A novel marker-free genome engineering approach in Cupriavidus necator, leveraging native partitioning systems for plasmid stability, enables sustainable recombinant protein production without antibiotic resistance genes.
Overview
This innovative solution focuses on marker-free genome engineering in Cupriavidus necator, a promising bacterial host for sustainable recombinant protein production. Utilizing tools developed by Technische Universitat Graz, this approach targets essential genes to create knockout strains. Plasmids are naturally maintained by strong native partitioning systems, eliminating the need for antibiotic resistance genes and aligning with industry trends to reduce antibiotic use. C. necator's ability to grow on diverse carbon sources makes it ideal for food and feed applications, providing an eco-friendly alternative for carbon reduction.
Technical specifications
- Marker-free knockout strains: Target essential genes in Cupriavidus necator without using antibiotic resistance markers.
- Native partitioning system: Ensures plasmid stability and reduces genetic drift, maintaining high copy numbers (20-30 copies/cell) without selective pressure.
- Versatile growth capabilities: Supports growth on sugars, waste streams, CO₂, and H₂, enabling sustainable production processes.
- Scalability: From small-scale (mL) to pilot-scale (up to 300 L) using gas fermenters, facilitating industrial-level recombinant protein production.
Technology readiness level
The technology is currently at TRL 3, with plans to advance to TRL 6 or higher through pilot-scale testing, focusing on plasmid stability and gene expression under various conditions, including heterotrophic and autotrophic growth.
About Technische Universität Graz
Graz University of Technology (TU Graz) is a STEM-focused public research university in Austria with a strong applied, industry-oriented profile. Its campuses sit within Graz’s manufacturing and mobility ecosystem, giving companies access to instrumented labs, large-scale testbeds, and prototyping spaces. Dedicated industry engagement teams coordinate contract research, sponsored theses, and tailored talent pipelines, while a regional science park and incubator co-located with the university support venture creation. Research is supported by competitive funding from Austrian national agencies and the European Union. A dedicated technology transfer office manages IP, licensing, and collaboration agreements.
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