基于SELDI―TOF―MS冷却滩羊肉微生物差异蛋白分析.docVIP

基于SELDI―TOF―MS冷却滩羊肉微生物差异蛋白分析 　　摘 要：以托盘密封包装的冷鲜滩羊肉为研究对象，基于组合蛋白芯片与表面增强激光解吸电离飞行时间质谱（surface enhanced laser desorption ionization time-of-flight mass spectrometry，SELDI-TOF-MS）平台对滩羊肉表面微生物蛋白进行分析，区分不同贮藏时间生物样本之间的差异。通过正交矫正偏最小二乘法判别分析判断模型第一主成分的得分值（VIP阈值1），并结合学生氏t检验的P值（阈值0.05）寻找差异性表达蛋白。结果表明：应用Biomarker Wizard软件分析找出差异蛋白峰132 个，其中75 个差异蛋白峰可根据质荷比经Swiss-Prot数据库检索出对应的蛋白质，其余57 个差异蛋白峰对应的为未知蛋白；从上述差异蛋白中未找到酶类的差异蛋白，但是找到了可能和滩羊肉菌体有关联的19 个菌体蛋白。SELDI-TOF-MS技术能够用于检测冷鲜滩羊肉不同贮藏阶段基因表达的各种蛋白质，从而发现大量具有价值的蛋白质标志分子。 　　关键词：滩羊肉；微生物；表面增强激光解吸电离飞行时间质谱；差异蛋白 　　Abstract： Microbial proteins on the surface of tray-packaged chilled Tan sheep meat were analyzed by surface enhanced laser desorption ionization time-of-flight mass spectrometry （SELDI-TOF-MS） and biological samples collected at different times during chilled storage were differentiated. The variable importance in the projection （VIP） value （threshold 1） of the first principal component of the orthogonal partial least squares-discriminant analysis （OPLS-DA） model was determined and combined with the P value （threshold 0.05） of student’s t test to find out the differentially expressed proteins. The results showed that 132 differential protein peaks were obtained by Biomarker Wizard software， 75 of which were identified by searching in the Swiss-Prot database based on their mass-to-charge ratio， and the remaining 57 were unknown proteins. These differential proteins did not include enzymes but included 19 proteins that might associated with bacteria on mutton meat. In summary， SELDI-TOF-MS allows the detection of various differentially expressed microbial proteins at different storage times of chilled Tan sheep meat and consequently the identification of a large number of valuable protein markers. 　　Keywords： chilled Tan sheep meat； microorganism； surface enhanced laser desorption ionization time-of-flight mass spectrometry （SELDI-TOF-MS）； differential protein 　　DOI：10.7506/rlyj1001-8123-201802006 　　中图分类号：TS251.1 文献标志码：A 文章编号：1001-8123（2018）02-003