When Biotechnology Meets Chemistry: Metabolic and Chemical Stable Isotope Labelling of RNA Building Blocks for the Modular Synthesis of (2H/13C/15N)-N1-Methylpseudouridine (m1Ψ) compounds

The development of processes for the utilisation of renewable resources is one of the main challenges of our society. Autotrophic fermentations, in which CO2 is converted into biomass, constitute a strategy towards sustainable carbon-cycle economy. In its cutting-edge research project, SI-MEPSI will combine biotechnological and chemical approaches to create an efficient route from CO2 fixation to the production of fine chemicals. In the first stage, SI-MEPSI will optimise H2/O2/CO2-based autotrophic fermentations with Ralstonia eutropha to develop an industrial production of stable-isotope-labelled (SI-labelled) RNA under non-explosive conditions. To achieve this objective, a thorough investigation into the dependence of the growth rate on physicochemical parameters, particularly the pressure, will be performed. Subsequently, the obtained SI-labelled RNA products will be employed as building blocks in a three-step, chemo-enzymatic synthesis of novel SI(2H/13C/15N)-labelled N1-methylpseudouridine (m1Ψ) compounds – important targets in the context of RNA stability studies and NMR structural analysis. Preparation of SI-labelled m1Ψ compounds with a range of site-specific isotopic patterns will rely on a modular methodology that introduces SI-labels into the target molecules at three different points. To maximise the efficiency of the research and facilitate knowledge transfer, the fermentation studies will take place at a biotechnology company in Germany, while the chemo-enzymatic synthesis will be carried out at a university in Austria. In addition, a dedicated training program within an already existing EU ITN will further increase the visibility of the applicant and reinforce his career prospects in the EU. Through targeted dissemination activities, SI-MEPSI will also enhance the competitiveness of sustainable industrial autotrophic processes in the European Biotechnology sector, create new job opportunities, and inspire young students to pursue a scientific career.