交联海藻酸钠磁性微球的制备及固定化胰蛋白酶研究

Study on Preparation of Magnetic Particle of Crosslinked Sodium Alginate and Immobilization of Trypsin

制备了交联海藻酸钠磁性微球,并以磁性微球为载体,戊二醛为交联剂,将胰蛋白酶固定化;利用透射电镜、粒度分析、红外分析对交联海藻酸钠磁性微球进行了表征;探讨给酶量、戊二醛浓度和pH值对固定化酶活性的影响;与自由酶比较,考察了固定酶的酶学性质。结果表明:交联海藻酸钠磁性微球是固定化胰蛋白酶的良好载体,固定化酶最适宜的条件是吸附时间12h,给酶量为100mg/0.1g磁性载体、交联剂戊二醛浓度为5%、溶液pH值为6,同时将酶固定化后,酶的稳定性和催化性能均有所提高。

The aim of this paper is to prepare magnetic particle of crosslinked sodium alginate and to apply it to immobilize trypsin. The magnetic particle of crosslinked sodium alginate are yielded by method of embedding on Fe3O4 particle. Trypsin is immobilized on magnetic particle of crosslinked sodium alginate through crosslinked reaction with glutaraldehyde. The magnetic particle of crosslinked sodium alginate are characterized by methods of TEM, particle size analysis and IR analysis. The effects of amount of enzyme given, concentration of glutaraldehyde and pH value in the process of preparation on the activity of magnetic immobilixed enzyme are studied. The characteristics of enzymology of the magnetic immobilized trypsin are discussed on the basis of being compared with free enzyme. Research reveals magnetic particle of crosslinked sodium alginate to be a good carrier of immobilized trypsin. The optimum conditions of immobilized trypsin are amount of enzyme given for 100mg/0. lg magnetic carrier, concentration of glutaraldehyde for 5% ,pH value for 6 and adsorption time for 12h.