Optimisation of induction conditions for a bacterial strain producing proinsulin aspart

Diabetes poses a serious threat to the health of people around world. Therefore, in 2021, World Health Organisation launched Global Compact, an initiative aimed at improving management and prevention diabetes. The rapid growth number diabetic patients has increased need for insulin. Rapid-acting human insulin analogues, including aspart, improve efficacy therapy. Methods aspart production include its biosynthesis proinsulin form Escherichia coli . However, yield recombinant protein largely depends on optimisation process. aim study was optimise induction conditions E. strain expressing through applying Design Experiment (DoE) approach enhance bacterial cell productivity. Materials methods. focused producing aspart. authors planned experiment using MODDE software reduced face-centred central composite design (CCF) enabling assessment factor interactions creation spaces. carried out fermentations 5 L Biostat® B bioreactor measured concentrations by capillary gel electrophoresis. results were analysed GraphPad Prism 6. Results. Using DoE approach, optimised producer Based data from response surface plots wet biomass concentration, specific productivity, volumetric as well plotted models, established spaces expression models demonstrated high predictive power reproducibility results. successfully validated process synthesis under conditions. productivity 3.06±0.16 g/L (conventional conditions) 4.93±0.80 (optimised conditions). Conclusions. achieved 60% increase may be used intensify industrial