Testing of A Suitable Procedure of Immobilization The Proteolytic Enzymes Into Silica Nanofibers As A Painless Enzyme Debridement
We used silica nanofibers as a substrate for covalent immobilization of proteolytic enzymes.Nanofibers with enzymes should serve as enzymatic debridement (removing of necrotic tissue by catalysis of proteolytic reaction - hydrolysis of the peptide bonds that link amino acids together in a polypeptide chain). The research was divided into two parts - the discovery of the most suitable ligand (cytotoxicity tests were carried out) and the discovery of the most suitable proteolytic enzyme (pH dependence of enzymes tests and enzyme stability tests were carried out).Already from earlier testing, the enzyme immobilization method with N-Hydroxysuccinimide ester formed from N-Hydroxysuccinimide and N- (3-Dimethylaminopropyl) -N′-ethylcarbodiimide hydrochloride as a very suitable method for stable immobilization of enzymes was demonstrated. However, this procedure was cytotoxic for tested fibroblasts. Thus, the method with N, N′-Disuccinimidyl carbonate is referred to as the best method for covalent immobilization of enzymes to silica nanofibers. The best of the tested enzymes are protease from Aspergillus oryzaeand trypsin from bovine pancreas.
Keywords - Enzymatic Debridement, Silica Nanofibers, Immobilization Of Protease, Necrotic Tissue