应用拉曼光谱对比分析德式乳杆菌保加利亚亚种ND02及其VBNC态细胞成分  

Comparative Analysis of Cellular Components of Lactobacillus delbrueckii subsp.bulgaricus ND02 in Normal and Viable but Non-culturable State by Raman Spectroscopy

作　　者：包秋华[1] 马学波 任艳 王丽娜[1] 张雨虹 代利霞 BAO Qiuhua;MA Xuebo;REN Yan;WANG Lina;ZHANG Yuhong;DAI Lixia(Key Laboratory of Dairy Biotechnology and Engineering,Ministry of Education,Key Laboratory of Dairy Products Processing,Ministry of Agriculture and Rural Affairs,Inner Mongolia Agricultural University,Hohhot 010018,China;School of Life Science and Technology,Inner Mongolia University of Science and Technology,Baotou 014016,China)

机构地区：[1]内蒙古农业大学乳品生物技术与工程教育部重点实验室,农业农村部奶制品加工重点实验室,内蒙古呼和浩特010018 [2]内蒙古科技大学生命科学与技术学院,内蒙古包头014016

出　　处：《食品科学》2022年第10期114-118,共5页Food Science

基　　金：国家自然科学基金地区基金项目(31860434);内蒙古自然科学基金项目(2019MS03051);财政部和农业农村部:国家现代农业产业技术体系项目。

摘　　要：采用扫描电子显微镜、单细胞拉曼光谱技术对比分析德氏乳杆菌保加利亚亚种ND02(Lactobacillus delbrueckii subsp.bulgaricus ND02)正常及其非可培养态(viable but non-culturable,VBNC)细胞的表观形态和细胞内部大分子化合物的变化。扫描电子显微镜结果显示,与正常细胞相比,德氏乳杆菌保加利亚亚种ND02的VBNC态细胞变短、变粗,且细胞表面发生褶皱;VBNC态细胞的拉曼光谱与正常细胞差异显著,主要集中在1673、1650、1459、1030、1005 cm^(-1)等处的12个峰,主要反映细胞内蛋白质、核酸等生物大分子变化,结果表明德氏乳杆菌保加利亚亚种ND02在诱导进入VBNC态时可能通过调整核酸、脂类和蛋白质适应不利环境的影响;研究结果也证实拉曼光谱技术可以在单细胞水平上对德氏乳杆菌保加利亚亚种ND02及其VBNC态细胞内大分子物质进行无损、无标记检测分析。In this study,scanning electron microscopy(SEM)and single-cell Raman spectroscopy(SCRS)were used to analyze the differences in the cellular morphology and intracellular macromolecular compounds of Lactobacillus delbrueckii subsp.bulgaricus ND02 in the normal and viable but non-cultivable(VBNC)states.The SEM results showed that the VBNC cells,with a wrinkled surface,were shorter and thicker than the normal ones.The Raman spectra of the VBNC cells were significantly different from those of the normal cells,mainly in terms of 12 peaks,such as 1673,1650,1459,1030,and 1005 cm^(-1),reflecting changes in intracellular biomacromolecules such as proteins and nucleic acids.It was shown that L.delbrueckii subsp.bulgaricus ND02 could adapt to adverse environments by regulating the metabolism of proteins,nucleic acids and lipids when induced to enter into VBNC state.The results also confirmed that single-cell Raman spectroscopy can allow non-destructive label-free detection of macromolecules inside normal and VBNC cells of L.delbrueckii subsp.bulgaricus ND02.

关 键 词：德氏乳杆菌保加利亚亚种ND02 活的非可培养态 诱导 单细胞拉曼光谱 细胞成分 

分 类 号：Q939.97[生物学—微生物学]