Along the lifetime of the Project, a considerable number of important results and developments has materialized, including:
• The soil bacterium Pseudomonas putida has been upgraded as an optimal chassis for hosting environmentally significant new to nature reactions. This has involved a plethora of genetic tools for genome editing, containment for the sake of biosafety, production of chromosome-less vesicles and vectors for inter-species genome shuffling and transplantation. This works showcases the power of Synthetic Biology to deliver new materials and activities for the sake of industrial and environmental catalysis.
• An unprecedented progress has been made re. engineering and expression of entirely artificial metalloenzymes by capitalizing on the unique structure and properties of streptavidin. This enabled incorporation of [Fe4S4]-clusters and thus designing and evolving artificial metalloenzymes (ArM) for the reduction of CO2 to alka/enes (Fischer-Tropschase). These developments open immense perspectives of valorization of CO2 into added-value molecules.
• Procedures to synthesize organo-silicon compounds, although still in small amounts, have been developed and their incorporation to bacterial metabolic networks or biomass attempted. The wealth of knowledge on xenobiology accumulated along the way will be invaluable for future endeavours aimed at generating life forms that incorporate in their biomass a non-biological element and thus expand the scope of possible biochemistries in live systems.
• An entirely new concept for automatic and programmable chemical synthesis stemmed from earlier attempts to code-back reactions to DNA. This involves capturing the chemical synthesis literature from manual operation to a fully described and universal Chemical Description Language (χDL) to be run automatically in the Chemical Processing Unit, or ChemPU. This technology opens new perspectives for production of complex chemical structures.
• Novel models to predict risks of introducing new-to-nature reactions in the Biosphere with Synthetic Biology agents have fore the first time being produced. Such models exposed the limitations and opportunities of large-scale environmental interventions, in particular the containment that the naturally-occurring ecosystems impose to the spreading of new agents. Such a knowledge should improve the perception of risks associated to spreading of SynBio agents.
• Despite the COVID impasse, a strong dissemination / communication program was unfolded during the Project, including artists in Residence, exhibitions, videos aimed at making the MADONNA research and proposal of large-scale, SynBo-based environmental interventions acceptable by the general public and regulatory bodies with the basic idea of fostering a much needed revision of our mutuality with the natural world.
