The usage of fossil resources leading to increasing atmospheric CO2 levels and global climate change should be rapidly replaced by implementing a circular economy. Circular bioeconomy converting sustainable substrates in moderately operating bioprocesses offers a plenitude of solutions. While synthetic biology provides a multitude of tools for strain engineering, their rapid use in hosts for optimal performance under industrial conditions is still challenging. Promising innovations are often trapped in the ‘valley-of-death’ as strain engineering faces a too complex space of putative manipulations. Novel approaches are needed to increase speed and success rate of strain and bioprocess engineering. The bio-intelligent approach, rigorously applied in BIOS, aims to accelerate and improve the conventional ‘design-build-test-learn’ (DBTL) cycle for strain and bioprocess engineering. Interdisciplinary collaboration will bridge microbiology, molecular biology, biochemical engineering with informatics, automation engineering, and mechanical engineering. Novel innovative metrics, biosensors, and bioactuators are developed for bi-directionally communication at biological-technical interfaces. Digital twins are created mimicking cellular and process levels. Integrating AI not only improves prediction quality but also enables hybrid learning, the key reason to increase speed and success rate in the novel bio-intelligent DBTL cycle (biDBTL). The power of biDBTL will be showcased by creating P. putida producer strains for terpenes, polyolefines, and methylacrylate. All are highly attractive products with a high potential for reducing anthropogenic greenhouse footprint. BIOS will open the door to a de-centralized, networked collaboration for strain and process engineering that efficiently links individual expertise for the sake of a symbiotic and rapid progress. BIOS also paves the way to de-centralized bio-manufacturing by implementing autonomous, self-controlled bioprocesses.

We aim to engineer the lifestyle of Pseudomonas putida to generate a tailored, re-factored chassis with highly attractive new-to-nature properties, thereby opening the door to the production of thus far non-accessible compounds. This industrially driven project capitalises on the outstanding metabolic endowment and stress tolerance capabilities of this versatile bacterium for the production of specialty and bulk chemicals. Specifically, we will build streamlined P. putida strains with improved ATP availability utilizing this power on demand, decoupled from growth. The well-characterized, streamlined and re-factored strain platform will offer easy-to-use plug-in opportunities for novel, DNA-encoded functions under the control of orthogonal regulatory systems. To this end, we will deploy a concerted approach of genome refactoring, model-driven circuit design, implementation of ATP control loops, structured modelling and metabolic engineering. By drawing on a starkly improved, growth-uncoupled ATP-biosynthetic machinery, empowered P. putida strains will be able to produce a) n-butanol and isobutanol and their challenging gaseous derivatives 1-butene (BE) and (iso-)butadiene (BDE) using a novel, new-to-nature route starting from glucose, as well as b) new active ingredients for crop protection, such as tabtoxin, a high-value, ß-lactam-based secondary metabolite with a huge potential as a new herbicide. The game-changing innovations brought in – in particular the uncoupling of ATP-synthesis and production from growth - will provide strong versatility, enhanced efficiency and efficacy to the production processes, thereby overcoming current bottlenecks, matching market needs and fostering high-level research growth and development.