摘要
目的 探讨转谷氨酰胺酶催化的交联对鸡蛋过敏原溶菌酶IgG和IgE结合能力的影响,为低敏鸡蛋制品加工提供理论依据和数据支撑。方法 首先利用离子交换层析分离制备溶菌酶纯品。将溶菌酶与转谷氨酰胺酶共孵育后,通过SDS-PAGE观察交联情况,利用兔抗溶菌酶特异性血清验证交联产物IgG结合能力的变化,利用鸡蛋过敏患者血清验证交联产物IgE结合能力,同时确定交联降低溶菌酶致敏性的最适反应条件。结果 转谷氨酰胺酶能够催化溶菌酶发生交联反应,在变换转谷氨酰胺酶添加量和交联时间后,能够形成不同分子量的交联产物。交联后的溶菌酶IgG结合能力显著降低,然而IgE结合能力仅在转谷氨酰胺酶与底物质量比为10 U/g、反应时间为6 h时较未加工溶菌酶降低,最低值下降32%。结论 转谷氨酰胺酶能够催化鸡蛋过敏原溶菌酶发生交联反应,然而交联后产物的IgE结合能力降低结果并不理想。
Objective To explore the effect of transglutaminase catalyzed cross-linking on the IgG and IgE binding ability of egg allergen lysozyme,so as to provide theoretical basis and data support for the processing of hypo-allergenic egg products.Methods First,ion exchange chromatography was used to separate and prepare pure lysozyme.After co-incubation of lysozyme with transglutaminase,its cross-linking was observed by SDS-PAGE,the change in the IgG binding capacity of the cross-linked product was verified by using rabbit anti-lysozyme specific serum,and the IgE binding capacity of the cross-linked product was verified by using the sera of egg allergy patients.At the same time,the optimal reaction conditions for cross-linking to reduce the allergenicity of lysozyme were determined.Results Transglutaminase could catalyze the cross-linking reaction of lysozyme,and the IgG binding ability of lysozyme after cross-linking was significantly reduced,but its IgE binding ability only changed at transglutaminase/substrate of 10 U/g,and at reaction time of 6 h.The lowest IgE binding capacity reduced about 32%.Conclusion Transglutaminase can catalyze the cross-linking reaction of egg allergen lysozyme,but the IgE binding ability of the cross-linked product was not reduced ideally.
作者
张晨晨
黎江东
周荣钦
麻小娟
Zhang Chenchen;Li Jiangdong;Zhou Rongqin;Ma Xiaojuan(School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China)
出处
《遵义医科大学学报》
2024年第1期18-24,共7页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:31660443)
遵义医科大学优秀青年人才项目(NO:18zy-009)
贵州省科技厅项目[NO:黔科合基础(2019)1348号]。
