超声波预处理对小清蛋白半乳糖糖基化特性的影响  

Effect of Ultrasound Pretreatment on Glycation Characteristics of Parvalbumin with Galactose

作　　者：姜晴 陈文美 邵艳红 涂宗财[1,2] 刘俊 JIANG Qing;CHEN Wenmei;SHAO Yanhong;TU Zongcai;LIU Jun(National R&D Center for Freshwater Fish Processing,College of Life Sciences,Jiangxi Normal University,Nanchang 330022,China;State Key Laboratory of Food Science and Resources,Nanchang University,Nanchang 330047,China)

机构地区：[1]江西师范大学生命科学学院,国家淡水鱼加工技术研发专业中心,江西南昌330022 [2]南昌大学食品科学与资源挖掘全国重点实验室,江西南昌330047

出　　处：《食品科学》2023年第23期21-26,共6页Food Science

基　　金：现代农业产业技术体系建设专项(CARS-45);国家自然科学基金地区科学基金项目(31960457,22368024);江西省自然科学基金面上项目(20224BAB205046);江西省技术创新引导类计划项目(20212AEI91001)。

摘　　要：糖基化是美拉德反应的初始阶段,在食品加工与贮藏中有着重要的作用。本实验采用超声波对鲢鱼小清蛋白进行预处理,随后将其与半乳糖进行糖基化反应,采用高效液相色谱联合尺寸排阻色谱,内源荧光、紫外吸收、同步荧光光谱以及高分辨质谱等技术对糖基化小清蛋白的多级结构、糖基化程度以及糖基化肽段和位点进行表征,探究超声波预处理对小清蛋白半乳糖糖基化特性的影响。结果表明:糖基化可明显增加小清蛋白分子质量,降低其游离氨基质量浓度、内源荧光及紫外吸收强度,并改变小清蛋白的三级结构,而超声波预处理进一步促进了这些变化。未经超声波处理的糖基化小清蛋白含有4个糖基化修饰位点(K46、K55、K65和K88),而超声波预处理结合糖基化后的小清蛋白糖基化修饰位点增加至6个(K46、K55、K65、K88、K97和K108)。综上,超声波预处理使蛋白质的结构展开,增加糖基化反应程度,改善蛋白质糖基化特性。Glycation is the initial stage of the Maillard reaction and plays an important role in food processing and storage.This study aimed to explore the effect of ultrasound pretreatment on the glycation characteristics of parvalbumin with galactose.The multilevel structure,glycation degree,glycated peptides and sites of glycated parvalbumin were characterized by high performance liquid chromatography(HPLC)combined with size exclusion chromatography(SEC),intrinsicfluorescence,ultraviolet absorption,synchronousfluorescence spectroscopy and high-resolution mass spectrometry(HR-MS).The results showed that glycation significantly increased the molecular mass,reduced the free amino content,intrinsicfluorescence and ultraviolet absorption intensity,and changed the tertiary structure of parvalbumin.This effect was further promoted by ultrasound pretreatment.The glycated parvalbumin without ultrasound pretreatment contained four glycation sites(K46,K55,K65 and K88),while the number of glycation sites increased to six(K46,K55,K65,K88,K97 and K108)after ultrasonic pretreatment combined with glycation.Therefore,ultrasound pretreatment can result in protein unfolding,increase the degree of glycation,and thereby improve the characteristics of protein glycation.

关 键 词：超声波 糖基化特性 小清蛋白 半乳糖 糖基化修饰位点 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]