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作 者:孟俊丽 吴敏[1] 丁稳 李颖[1] 张进 倪恨美[1]
出 处:《Journal of Southeast University(English Edition)》2014年第4期526-530,共5页东南大学学报(英文版)
基 金:The National Natural Science Foundation of China(No.21005016);the Foundation of Educational Commission of Jiangsu Province(No.JHB2011-2)
摘 要:In order to improve the substrate diffusion properties and stability of an immobilized enzyme alginate microgels modified with TiO2 nanoparticles were employed as the enzyme immobilizing support.Ionotropic gelation was applied for the preparation of hybrid gels while Ca2+ Ce3+ Ni2+Cu2+and Fe3+were employed as the crosslinkers.Papain was selected as the model enzyme. UV-Vis spectroscopy was employed to investigate the activity of papain to evaluate kinetics and stability.Analysis results show that the highest affinity the lowest Michaelis-Menten constant Km =11.0 mg/mL and the highest stability are obtained when using Cu2+as the crosslinker.The effect of the mass ratio of TiO2 to papain on the stability and leakage of papain is also investigated and the results show that 10∶1 TiO2∶papain is optimal because the proper use of TiO2 can reduce enzyme leakage and ensure enzyme stability.Preparing Cu/alginate/TiO2 hybrid gels via ionotropic gelation can provide a satisfactory diffusion capability and enzyme stability.为了改善海藻酸钠固定化酶微球的扩散性与酶活稳定性,以纳米二氧化钛修饰海藻酸为固定化酶载体,Ca2+,Ce3+,Ni2+,Cu2+和Fe3+为杂化凝胶的阳离子交联剂,木瓜蛋白酶为模型酶,制备固定化酶微球.通过UV-Vis光谱检测酶活,考察了5种交联剂对固定化酶的动力学和稳定性的影响,结果表明:当Cu2+为交联剂时,固定化酶具有最高的亲和性、最低的米氏常数(Km=11.0 mg/m L)和最高的稳定性.研究了TiO 2与木瓜蛋白酶的质量比对固定化酶稳定性以及酶蛋白泄漏的影响,结果显示纳米二氧化钛与木瓜蛋白酶的质量比为10∶1时,制备的固定化酶微球性能最好,因为二氧化钛能有效阻止固定化酶的泄漏.通过离子胶凝制备Cu/海藻酸/TiO 2杂化凝胶可获得较好的扩散能力和酶活稳定性.
关 键 词:cationic crosslinker diffusion hybrid gel ionotropic gelation Michaelis-Menten constant titanium dioxide modified alginate
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