Trypsin Encapsulation within Pectin-Poly(diallyldimethylammonium Chloride) Complex Coacervates

Active substances like pharmaceuticals and enzymes can be shielded from a variety of unfavourable environmental conditions, including high pH, organic solvents, and chaotic agents, by employing complex coacervation as an encapsulation strategy. One of the goals of this research is to form complex coacervate droplets using pectin, a carbohydrate present in plant cell walls, and poly(diallyldimethylammonium chloride), a synthetic homopolymer. Electrostatic interactions between the positive charges in PDADMAC and the negative charges in pectin are the main factors for coacervation to assemble. Encapsulation of the trypsin enzyme within complex coacervates made of PDADMAC and pectin was examined as a function of mixing order of these polyelectrolytes and at various salt concentrations to learn more about protein encapsulation within complex coacervates. There were three different mixing orders used. We examined the effect of ionic strength on pectin-PDADMAC system by turbidimetric titrations at six different salt concentrations. Light microscopy allowed us to observe the formation of coacervate microdroplets at these salt concentrations. In order to determine the degree of ionization of pectin at the optimal pH for trypsin, which is 7.5, potentiometric titration studies were carried out on pectin. The most efficient mixing order is chosen for the subsequent experiments. The impact on enzyme encapsulation was also investigated by varying the amounts of trypsin and polyelectrolytes. Enzyme activity was assessed following the selection of the most efficient encapsulation technique. Circular dichroism studies were used to determine if the secondary structure of trypsin altered during encapsulation.