摘要
将溶菌酶与卵黏蛋白相互作用,以溶菌酶与卵黏蛋白为对照,研究溶菌酶与卵黏蛋白在不同反应时间(10、20、30、40、50、60 min),温度(5、15、25、35、45℃),pH(7.0、7.5、8.0、8.5、9.0),质量比(0.50∶1.00、0.75∶1.00、1.00∶1.00、1.00∶0.75、1.00∶0.50)下相互作用的浊度、表观黏度的变化;通过SDS-PAGE凝胶电泳、傅里叶红外光谱、低场核磁共振技术、扫描电镜对复合物的类型、作用力及结构等进行表征。结果表明:在反应时间为40min、反应温度为25℃、pH为8.5、溶菌酶与卵黏蛋白的质量比为0.75∶1.00时,溶菌酶与卵黏蛋白相互作用最强;相较于卵黏蛋白和溶菌酶,复合物中α-螺旋和β-转角占比增加,β-折叠和无规则卷曲占比减少;互作后其结合水含量增加,自由水含量有所降低,疏水性高于溶菌酶,而低于卵黏蛋白;卵黏蛋白与溶菌酶互作包括氢键和二硫键结合及存在静电相互作用、表面疏水作用等分子间作用力;互作后微观结构发生变化,呈现出絮状结构。表明蛋白质间相互作用会导致其结构和聚集状态发生较大变化,产生不溶性复合物,可能与蛋清浓蛋白的黏弹性有关。
This study aimed to investigate the interaction mechanism between lysozyme and ovomucin.With lysozyme and ovomucin as controls,the turbidity and apparent viscosity of the interaction between lysozyme and ovomucin were studied under six reaction time(10,20,30,40,50,60 min),five temperatures(5,15,25,35,45℃),five pH(7.0,7.5,8.0,8.5,9.0),and five mixing ratios(0.50∶1.00,0.75∶1.00,1.00∶1.00,1.00∶0.75,1.00∶0.50).The acting force and structure of aggregates were characterized by SDS-PAGE gel electrophoresis,Fourier transform infrared spectroscopy,low-field nuclear magnetic resonance and scanning electron microscope.The results showed that the conditions of best interaction between lysozyme and ovomucin were incubation time of 40 min,reaction 25℃,pH of 8.5,with the ratio of 0.75∶1.00.Compared with ovomucin and lysozyme alone,the complex owned moreα-helix andβ-turn,and lessβ-pleated sheet and irregular coil.After interaction,the bound water content increased,and the free water decreased.The hydrophobicity of complex was higher than that of lysozyme,but lower than that of ovomucin.There were electrostatic interaction,hydrophobic interaction,hydrogen bond,disulfide bond and other intermolecular forces in the interaction between ovalbumin and lysozyme.After interaction,the microstructure changed and then flocculent structure was showed.It indicated that the protein-protein interactions would lead to great changes in their structure and aggregation state and produced insoluble complex,which might be related to the viscoelasticity of egg white protein.
作者
刘丽莉
张孟军
程伟伟
杨晓盼
徐宝成
肖枫
LIU Lili;ZHANG Mengjun;CHENG Weiwei;YANG Xiaopan;XU Baocheng;XIAO Feng(College of Food&Bioengineering,Henan University of Science and Technology,Luoyang,Henan 471023,China;National Teaching Demonstration Center for Food Processing and Safety,Luoyang,Henan 471023,China;Henan Engineering Technology Research Center for Food Raw Materials,Luoyang,Henan 471023,China;Research and Utilization of Functional Food Resources Scientific and Technological Innovation Team of Henan Provincial Departmentof Education,Luoyang,Henan 471023,China)
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2022年第6期672-678,729,共8页
Journal of Hunan Agricultural University(Natural Sciences)
基金
国家自然科学基金项目(U1704114)
2021年洛阳市社会发展专项(2101021A)。
关键词
蛋清
溶菌酶
卵黏蛋白
相互作用
浊度
表观黏度
结构
egg white
lysozyme
ovomucin
interaction
turbidity
apparent viscosity
structure
