基质辅助激光解析电离飞行时间质谱法分析天然牦牛酸奶中的德氏乳杆菌
目的 探索基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry, MALDI-TOF MS)在牦牛酸奶中德氏乳杆菌检测中的应用。方法 采用MALDI-TOF MS对分离自天然牦牛酸奶中的德氏乳杆菌进行分析研究。针对10株分离自牦牛酸奶经16S rDNA鉴定的德氏乳杆菌进行质谱分析, 根据乳酸菌的特异性特征, 设计6种微生物破碎方法和5种微生物蛋白质抽提方法进行正交组合实验, 在同一质谱条件下采用加权计算及均方根误差分析的获得变异较低稳定性高的处理方法。在最佳处理组合下采用SARAMIS Premium对德氏乳杆菌进行相关分析。结果 实验结果表明在液氮、超声、甲酸处理下德氏乳杆菌在基质解析电离中可产生稳定且丰富的蛋白质峰组, 且相关分析较离散, 但实验结果可为菌种差异性分析提供理论帮助。结论 MALDI-TOF MS 对天然牦牛酸奶中德氏乳杆菌特征性分析具有较强应用价值。
Objective To explore the application of matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF MS) in the detection of Lactobacillus delbrueckii isolated from natural yak yogurt. Methods MALDI-TOF-MS was used to research the Lactobacillus delbrueckii isolated from natural yak yogurt. Ten strains of Lactobacillus delbrueckii isolated from yak yoghurt were identified by 16S rDNA and analyzed by mass spectrometry. According to the specific characteristics of lactic acid bacteria, 6 kinds of microbial crushing methods and 5 kinds of microbial protein extraction methods were designed for orthogonal combination experiment. Under the same mass spectrum condition, weighted calculation and root mean square error analysis were used to obtain the treatment method with low variation and high stability. In addition, SARAMIS Premium was used for the Lactobacillus delbrueckii correlation analysis under optimal treatment combination. Results The experimental results showed that Lactobacillus delbrueckii could produce stable and abundant protein peaks in the matrix analytical ionization under liquid nitrogen, ultrasound and formic acid treatment, and the correlation analysis was relatively discrete, but the experimental results could provide theoretical help for the analysis of the difference of strains. Conclusion MALDI-TOF MS has strong application value in the characteristic analysis of Lactobacillus delbrueckii in natural yak yoghurt.
标题：基质辅助激光解析电离飞行时间质谱法分析天然牦牛酸奶中的德氏乳杆菌
英文标题：Analysis of Lactobacillus delbrueckii isolated from natural yak yogurt by matrix-assisted laser desorption/ionization time of flight mass spectrometry
作者：
魏超 四川省农科院分析测试中心
代晓航 四川省农科院分析测试中心
郭灵安 四川省农科院分析测试中心
刘炜 四川省农科院分析测试中心
中文关键词:基质辅助激光解析电离飞行时间质谱法,德氏乳杆菌,牦牛酸奶,乳酸菌,标识峰,
英文关键词:matrix-assisted laser desorption/ionization time of flight mass spectrometry,Lactobacillus delbrueckii,yak yogurt,lactic acid bacteria,identify peak,
发表日期：2019-05-02
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