The BIOS project has advanced both the scientific basis and the technical toolbox for bio-intelligent biomanufacturing. On the modelling side, the consortium has built detailed “digital twins” of Pseudomonas putida that combine mechanistic descriptions of metabolism and gene expression with data-driven models. These twins can predict how genetic changes and process conditions influence cell performance and product formation, providing a much stronger basis for rational strain design and model-guided process optimisation.
Experimentally, BIOS has established an integrated platform for building and testing strains. This includes a modern genome engineering toolkit (CRISPR-based control, efficient DNA insertion and recombineering), high-throughput electroporation hardware for rapid transformation in microtiter plates, and optimised cell-free protein synthesis protocols for fast part screening. In parallel, robust fluorescent reporter systems and automated RNA-sequencing workflows have been implemented to quantify gene expression and stress responses in a systematic, reproducible way and to feed these data directly back into the design–build–test–learn cycle.
These capabilities have already led to concrete biological and process advances. BIOS has identified the main stress factors that limit terpene and C1-based production in P. putida and uncovered hidden loss of production capacity over time in lycopene strains, guiding new engineering strategies. For methacrylate esters and other target molecules, new process concepts such as in situ product removal have delivered large increases in titres and much simpler downstream processing. Finally, life-cycle and techno-economic assessments are being used to benchmark these new routes against conventional production, ensuring that the technical innovations are aligned with environmental performance and future industrial viability.
