Development and biotechnological applications of multifunctional and biomimetc proprotei̇n convertases (PPC)

In the scenario of absolute insulin deficiency resulting from autoimmune damage to pancreatic ß-cells, patients are compelled to administer exogenous insulin that mimics physiological insulin secretion for achieving optimal glycemic control. This study delves into the methodology of producing recombinant insulin as a biomimetic drug, wherein a mimic proprotein convertase enzyme catalyzing the transformation of recombinant proinsulin into mature insulin has been synthesized, and corresponding activity studies have been conducted. Within this framework, a mimic proprotein convertase enzyme was created by introducing methacrylate functional groups to histidine, aspartic acid, and serine amino acids in the active region of the proprotein convertase enzyme. This was achieved through molecular imprinting technology using the Pyr-Arg-Thr-Lys-Arg-MCA molecule as a template, which has not been previously documented in the literature. The activity results indicated a Vmax of 6.24 μmol/min and a Km of 3.23. In the 8th repetition of the synthesized mimic proprotein convertase enzyme, only a 14% loss in activity was observed. This not only addresses efficiency losses in the insulin production process by combining the A and B chains that constitute insulin but also presents an approach to reduce enzyme costs and purification steps in the enzymatic proinsulin-insulin transformation.