超速离心和酸沉淀制备牛乳清的蛋白质组学研究

doi:10.16431/j.cnki.1671-7236.2019.08.002

《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (8): 2203-2210.doi: 10.16431/j.cnki.1671-7236.2019.08.002

超速离心和酸沉淀制备牛乳清的蛋白质组学研究

齐云霞¹, 赵小伟¹, 黄冬维¹, 蔡东东², 杨永新¹, 程广龙¹, 赵辉玲¹

1. 安徽省农业科学院畜牧兽医研究所, 畜禽产品安全工程安徽省重点实验室, 合肥 230031;
2. 安徽省兽药饲料监察所, 合肥 230001

收稿日期:2018-12-04 出版日期:2019-08-20 发布日期:2019-08-17
通讯作者: 程广龙 E-mail:cgl0126@126.com
作者简介:齐云霞(1981-),女,山东曲阜人,博士,研究方向:分子营养,E-mail:qiyx09@163.com
基金资助:
公益性行业（农业）科研专项课题"生鲜乳质量特征评价技术研究"（201403071-3）；生鲜乳特质性营养品质评价与关键控制点评估（GJFP201700805）；安徽省牛羊产业技术体系

Proteomics Study on Preparation of Bovine Whey by Ultracentrifugation and Acid Precipitation

QI Yunxia¹, ZHAO Xiaowei¹, HUANG Dongwei¹, CAI Dongdong², YANG Yongxin¹, CHENG Guanglong¹, ZHAO Huiling¹

1. Anhui Key Laboratory of Livestock and Poultry Product Safety Engineering, Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agricultural Science, Hefei 230031, China;
2. Anhui Provincal Institute of Animal Drug and Feed Supervision, Hefei 230001, China

Received:2018-12-04 Online:2019-08-20 Published:2019-08-17

摘要/Abstract

摘要：

本研究旨在对超速离心法和酸沉淀法鉴定的乳清蛋白进行分析，并比较不同样品制备方法对乳清蛋白组鉴定结果的影响。采集牛奶样品去除脂肪，利用超速离心和酸沉淀法制备乳清蛋白，液相色谱串联质谱分析结合数据库搜索鉴定，比较两种方法制备的乳清蛋白的差异，分析了差异蛋白参与的生物过程、细胞组成和分子功能等。聚类分析和主成分分析展示了两种样品制备方法的差异蛋白，超速离心方法鉴定的表达量增加的蛋白有53个，包括β-酪蛋白、糖基化依赖细胞黏附分子1和多聚免疫球蛋白受体等；酸沉淀方法鉴定的表达量增加的蛋白有21个，包括CD9抗原、β2-微球蛋白和β-乳球蛋白等。将两种方法获得的差异蛋白根据蛋白注释分类发现，超速离心法中高表达的蛋白主要位于胞外区域、囊泡、内膜系统、内质网和高尔基体等，涉及蛋白结合、酶调节活性、受体结合和抗氧化活性等；酸沉淀法中高表达的蛋白主要位于胞外区域和囊泡等，涉及结合、核酸酶活性和离子结合等。结果提示，两种方法鉴定的蛋白质组表达模式存在差异，乳清蛋白样品制备方法的结合可以获得更多的鉴定蛋白，为乳清蛋白组学研究中样品制备方法的选择提供参考依据。

关键词: 牛奶; 乳清蛋白; 超速离心; 酸沉淀

Abstract:

This study was aimed to analyze whey proteins identified by ultracentrifugation and acid precipitation, and compare the effects of different sample preparation methods on the identification results of whey proteome.Milk samples were collected to remove fat.Whey protein was prepared by ultracentrifugation and acid precipitation, the difference between two methods was compared by LC-MS and database search, the biological processes, cell composition and molecular functions of different proteins were analyzed.Cluster analysis and principal component analysis showed that 53 proteins with increased expression levels were identified by ultracentrifugation, including β-casein, glycosylation-dependent cell adhesion molecule 1 and polyimmunoglobulin receptors;21 proteins with increased expression levels were identified by acid precipitation method, including CD9 antigen, β2-microglobulin and β-lactoglobulin.According to annotation classification, proteins highly expressed in ultracentrifugation were mainly located in extracellular regions, vesicles, intimal system, endoplasmic reticulum and Golgi apparatus, involving protein binding, enzyme-modulating activity, receptor binding and resistance;Proteins highly expressed in acid precipitation were also mainly located in extracellular regions and vesicles, involving binding, nuclease activity, and ionic binding.Based on the above analysis results, the proteome profiles identified by two methods were different, and the combination of two methods could obtain more identified proteins.This study could provide a basis for the selection of sample preparation methods in whey proteomics research.

Key words: milk; whey protein; ultracentrifugation; acid precipitation

中图分类号:

TQ937

齐云霞, 赵小伟, 黄冬维, 蔡东东, 杨永新, 程广龙, 赵辉玲. 超速离心和酸沉淀制备牛乳清的蛋白质组学研究[J]. 《中国畜牧兽医》, 2019, 46(8): 2203-2210.

QI Yunxia, ZHAO Xiaowei, HUANG Dongwei, CAI Dongdong, YANG Yongxin, CHENG Guanglong, ZHAO Huiling. Proteomics Study on Preparation of Bovine Whey by Ultracentrifugation and Acid Precipitation[J]. , 2019, 46(8): 2203-2210.

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超速离心和酸沉淀制备牛乳清的蛋白质组学研究

Proteomics Study on Preparation of Bovine Whey by Ultracentrifugation and Acid Precipitation

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