Polyclonal antibodies mediated immobilization of a peroxidase from ammonium sulphate fractionated bitter gourd (Momordica charantia) proteins

Aiman Fatima, Qayyum Husain
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引用次数: 13

Abstract

Polyclonal antibody bound Sepharose 4B support has been exploited for the immobilization of bitter gourd peroxidase directly from ammonium sulphate precipitated proteins. Immunoaffinity immobilized bitter gourd peroxidase exhibited high yield of immobilization. IgG-Sepharose 4B bound bitter gourd peroxidase showed a higher stability against heat, chaotropic agents (urea and guanidinium chloride), detergents (cetyl trimethyl ammonium bromide and Surf Excel), proteolytic enzyme (trypsin) and water-miscible organic solvents (propanol, THF and dioxane). The activity of immobilized bitter gourd peroxidase was significantly enhanced in the presence of cetyl trimethyl ammonium bromide and after treatment with trypsin as compared to soluble enzyme.

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多克隆抗体介导的硫酸铵分离苦瓜蛋白过氧化物酶的固定化
利用多克隆抗体结合Sepharose 4B载体,利用硫酸铵沉淀蛋白直接固定化苦瓜过氧化物酶。免疫亲和固定化苦瓜过氧化物酶具有较高的固定化产量。IgG-Sepharose 4B结合的苦瓜过氧化物酶对高温、嗜热剂(尿素和氯化胍)、洗涤剂(十六烷基三甲基溴化铵和Surf Excel)、蛋白水解酶(胰蛋白酶)和与水混溶的有机溶剂(丙醇、四氢呋喃和二氧六环)具有较高的稳定性。在十六烷基三甲基溴化铵和胰蛋白酶处理下,固定化苦瓜过氧化物酶活性显著高于可溶性酶。
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