基于MALDI-TOF-MS技术筛选及鉴定大曲中芽孢杆菌  

Screening and identification of Bacillus in Daqu based on MALDI-TOF-MS technology

作　　者：李丹 张文学 LI Dan;ZHANG Wenxue(School of Liquor-Brewing Engineering,Sichuan University Jinjiang College,Meishan 620860,China;College of Biomass Science and Engineering,Sichuan University,Chengdu 610065,China)

机构地区：[1]四川大学锦江学院白酒学院,四川眉山620860 [2]四川大学轻工科学与工程学院,四川成都610065

出　　处：《中国酿造》2023年第7期121-125,共5页China Brewing

基　　金：国家重点研发计划(2018YFE0127400);四川省教育厅科研项目(18ZB0366)。

摘　　要：该研究对基质辅助激光解吸电离-飞行时间-质谱(MALDI-TOF-MS)技术体系进行改进与优化,创新性地引入浓香型大曲中芽孢杆菌(Bacillus)的筛选及鉴定,并结合分子生物学技术对该技术鉴定结果进行验证。结果表明,采用传统微生物培养分离法从大曲中分离得到126株菌株,其中125株通过MALDI-TOF-MS技术筛选鉴定获得9.0以上可信度分值,其中目标菌株枯草芽孢杆菌(Bacillus subtilis)及地衣芽孢杆菌(Bacillus licheniformis)的鉴定结果与分子生物学鉴定结果一致,说明该技术历经5~6 d可以准确地完成大曲样品中目标菌株的筛选鉴定,比传统微生物筛选鉴定方法至少节约1~2周时间。因此,基于MALDI-TOF-MS技术参与复杂样品中目标菌株的筛选与鉴定是一条高效的、可行的路径。The technology of matrix assisted laser desorption ionization-time of flight-mass spectrometry(MALDI-TOF-MS)was improved and optimized,and introduced into the screening and identification of Bacillus in strong-flavor(Nongxaingxing)Daqu,which was verified by molecular biological technique.The results showed that 126 strains were isolated from Daqu by traditional microbial culture and separation method,and 125 strains of them were screened and identified by MALDI-TOF-MS technique with a confidence score of more than 9.0.Among them,the identification results of target strains Bacillus subtilis and Bacillus licheniformis were in accordance with the results detected by molecular biology,which suggested that the screening and identification of target strains in Daqu samples could be completed accurately in 5-6 d by using MALDI-TOF-MS technology,saving at least 1-2 weeks compared with the traditional methods of microbial screening and identification.Therefore,the screening and the identification of target strains in complex samples based on MALDI-TOF-MS technology were an efficient and feasible approach.

关 键 词：基质辅助激光解吸电离飞行时间质谱 大曲 芽孢杆菌 蛋白指纹图谱 筛选 鉴定 

分 类 号：TS261.1[轻工技术与工程—发酵工程]