| Autor: |
Abdel-Mageed HM; a Molecular Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre , Cairo , Egypt.; b Faculty of Pharmaceutical Sciences and Pharmaceutical Industries, Future University in Egypt (FUE) , Cairo , Egypt., Radwan RA; c Biochemistry and Biotechnology Department, Faculty of Pharmacy and Drug Technology, Heliopolis University , Cairo , Egypt., AbuelEzz NZ; d Biochemistry Department, College of Pharmaceutical Sciences & Drug Manufacturing, Misr University for Science and Technology , Cairo , Egypt., Nasser HA; e Microbiology and Public Health Department, Faculty of Pharmacy and Drug Technology, Heliopolis University , Egypt., El Shamy AA; e Microbiology and Public Health Department, Faculty of Pharmacy and Drug Technology, Heliopolis University , Egypt., Abdelnaby RM; f Pharmaceutical Chemistry Department, Faculty of Pharmacy and Drug Technology, Heliopolis University , Egypt., El Gohary NA; g Pharmaceutical Chemistry Department, Faculty of Pharmacy and Biotechnology, German University in Cairo , Cairo , Egypt. |
| Abstrakt: |
Enzymes are powerful versatile biocatalysts, however, industrial application of enzymes is usually hampered by their susceptibility. Bio-inspired Eudragit-α-amylase conjugate (E-AC) was proposed as a biocatalyst for various pharmaceutical and industrial applications. In this study, α -Amylase (E.C. 3.2.1.1) was immobilized by covalent conjugation to Eudragit L-100 under mild conditions. The effect of polymer, carbodiimide and enzyme concentrations on optimization of (E-AC) was investigated. In addition, characterization of the free α -Amylase and E-AC with regard to pH, temperature, kinetic parameters, reusability and operational and storage conditions was carried out. Results showed a shift of the optimum pH of E-AC towards the alkaline side whereas, E-AC exhibited higher thermal stability at all tested temperatures. The kinetic parameters, K m values were 2.87 mg/ml and 3.15 mg/ml and V max values were 8.35 mg/ml/min and 8.98 mg/ml/min for free and E-AC, respectively. E-AC retained 85% of the initial activity after five consecutive amylolytic cycles, thus emphasizing its powerful potentials. Operational storage and thermal stability were highly improved as well for E-AC conjugate with an 11.6 stabilization factor in comparison to the free α-amylase. In this study, Eudragit L-100 polymer was successfully used as smart immobilization support to create a reversibly soluble-insoluble enzyme biocatalyst to enforce and extend biotechnological applications of α-amylase in the pharmaceutical industry. |
