Escherichia coli konak organizmada GLP-1 analoğunun rekombinant üretimi

Diabetes is the most serious metabolic disorder correlated with obesity, hypertension and cardiovascular conditions. High prevalence of Type II Diabetes Mellitus (T2DM) indicates the need for new medication development. In developing therapeutics, higher efficiency and fewer adverse effect features are targeted primarily. Recombinant protein-based biotechnological drug molecules have been developed and used for the treatment of T2DM. Especially, GLP-1 analogues are known by their self-limiting mechanism and insulinotropic effect. In this study, a novel GLP-1 analogue with increased stability and efficiency is produced using recombinant E. coli. The expression plasmid was constructed and confirmed by restriction digestion and whole plasmid sequencing. Then, itwas transformed into various E. coli strains followed by optimized lysis, growth and expression conditions to maximize the yield of the GLP-1 analogue. Various parameters such as pre-induction time, induction point, induction IPTG concentration and post-induction temperature were tested for the succesfull expression with maximum yield. Consequently, it was achieved that E. coli BL21(DE3) as strain, 0.2 mM IPTG induction at OD600nm of 0.6 and 18 °C overnight post-induction growth was the most promising conditions. Under these conditions, the GLP-1 analogue was obtained in the insoluble fraction. Following protein analysis and purification, quantification was performed and the highest titer of GLP-1 analogue was measured as 626 µg/ml. As future prospect, using another host organism and changing growth conditions can provide obtaining target protein in the soluble form. Keywords: T2DM, GLP-1 analogue, recombinant DNA technology, protein expression, E. coli