Immobilization of trypsin in organic and aqueous media for enzymatic peptide synthesis and hydrolysis reactions

摘要: Immobilization of enzymes onto different carriers increases enzyme’s stability and reusability within biotechnological pharmaceutical applications. However, some immobilization techniques are associated with loss enzymatic specificity and/or activity. Possible reasons for this mass transport limitations or structural changes. For reason an method must be selected depending on immobilisate’s demands. In work media were compared towards the synthetic hydrolytic activities immobilized trypsin as model enzyme magnetic micro-particles. Porcine was carried out in organic aqueous microparticles. The conditions solvent optimized a peptide synthesis reaction. highest carrier activity achieved at 1 % water (v/v) dioxane. resulting immobilizate could used over ten cycles retention 90 % reaction 80 % ethanol hydrolysis 87 % buffered solution. Further, applied reactions comparison to varying protein input. dioxane showed higher coupling yield by factor 2 maximum 19.2 % immobilization. displayed value 20.4 % while 38.5 %. Comparing specific yields tested methods revealed values 5.2 % 100 % 33.3 % 87.5 % method, respectively. By immobilizing dioxane, beneficial effect resilience medium shown. results indicate substantial potential micro-aqueous protease preservation during step. These may interest mild high selectivity industrial drug production.