Immobilization of serine protease subtilisin carlsberg on different carriers, characterization and kinetic properties in hydrolysis and transesterification reactions of serine protease subtilisin carlsberg

Subtilisin Carslberg (SC) is an enzyme belonging to the alkali serine endoprotease family obtained by fermentation from Bacillus licheniformis and hydrolyzes natural or denatured proteins under alkaline conditions. Subtilisin Carslberg consists of a single polypeptide chain that does not contain disulfide bonds. In addition, the isoelectric point is 9.4 and it can show its activity in some organic solvents. Besides its proteolytic activity, it also has esterase activity. In this study, Subtilisin Carlsberg was immobilized on magnetic chitosan (MK), perlite and β-cyclodextrin modified perlite (Perlit-CD) using two different crosslinkers: glutaraldehyde and genipin. The hydrolysis and transesterification activity of immobilized Subtilisin Carlsberg has been extensively investigated. The structure and surface morphology of prepared carriers were characterized by SEM, TGA and EDX. The optimum temperature and pH value of free and immobilized Subtilisin Carlsberg activity were found to be 50oC and pH 8.0, respectively. It was similar to the free enzyme at optimum pH and temperature for the activity of Subtilisin Carlsberg immobilized on genipin and glutaraldehyde activated carriers. With enzymatic hair removal as an application of the hydrolysis activity of Subtilisin Carlsberg, a softer and smoother skin surface was obtained compared to the chemical method. In studies on the transesterification activity of Subtilisin Carlsberg, the optimum temperature was determined as 50 oC. In addition, the effect of metal ion and salt addition on the transesterification activity was investigated.