| Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus |
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| 作者姓名： | Ming-ming CHEN Ai-li LI Mao-cheng SUN Zhen FENG Xiang-chen MENG Ying WANG |
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| 作者单位： | [1]MOE Key Laboratory of Dairy Science, College of Food Science, Northeast Agricultural University, Harbin 150030, China [2]School of Public Health, Jilin Medical College, Jilin 132013, China |
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| 基金项目： | supported by the Open Research Fund for the MOE Key Laboratory of Dairy Science（No.2012KLDSOF-07）; the National Natural Science Foundation of China（No.31000808）; the Synergetic Innovation Center of Food Safety and Nutrition; the Open Research Fund of Northeast Agricultural University,China |
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| 摘 要： | 研究目的：为保加利亚乳杆菌的代谢组学研究提供一种简单有效的淬灭方案。创新要点：采用适当的淬灭技术是获得微生物真实代谢物组数据的必要条件。本研究首次建立了适用于保加利亚乳杆菌代谢组学研究的淬灭方法,有助于完善乳酸菌代谢的生理学和遗传学概貌。研究方法：分别应用-40°C的60%甲醇/水、80%甲醇/水和80%甲醇/甘油三种方式淬灭保加利亚乳杆菌。利用光密度（OD）回收率实验、流式细胞术和气质联用（GC-MS）分析菌体细胞完整性和细胞内外代谢物,以及主成分分析法（PCA）和正交偏最小二乘法-判别分析法（OPLS-DA）分析代谢物泄露程度。重要结论：80%冷甲醇/水更适用于淬灭保加利亚乳杆菌,能有效减少代谢物泄露程度和增加胞内代谢物水平。
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| 关 键 词： | 代谢组学 淬灭处理 保加利亚乳杆菌 泄露 |
| 收稿时间： | 2013 May 24 |
Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus |
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| Ming-ming CHEN,Ai-li LI,Mao-cheng SUN,Zhen FENG,Xiang-chen MENG,Ying WANG.Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus[J].Journal of Zhejiang University Science,2014,15(4):333-342. |
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| Authors: | Ming-ming Chen Ai-li Li Mao-cheng Sun Zhen Feng Xiang-chen Meng Ying Wang |
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| Institution: | 1. MOE Key Laboratory of Dairy Science, College of Food Science, Northeast Agricultural University, Harbin, 150030, China
2. School of Public Health, Jilin Medical College, Jilin, 132013, China
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| Abstract: | This study proposed a quenching protocol for metabolite analysis of Lactobacillus delbrueckii subsp. bulgaricus. Microbial cells were quenched with 60% methanol/water, 80% methanol/glycerol, or 80% methanol/water. The effect of the quenching process was assessed by the optical density (OD)-based method, flow cytometry, and gas chromatography-mass spectrometry (GC-MS). The principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were employed for metabolite identification. The results indicated that quenching with 80% methanol/water solution led to less damage to the L. bulgaricus cells, characterized by the lower relative fraction of prodium iodide (PI)-labeled cells and the higher OD recovery ratio. Through GC-MS analysis, higher levels of intracellular metabolites (including focal glutamic acid, aspartic acid, alanine, and AMP) and a lower leakage rate were detected in the sample quenched with 80% methanol/water compared with the others. In conclusion, we suggested a higher concentration of cold methanol quenching for L. bulgaricus metabolomics due to its decreasing metabolite leakage. |
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| Keywords: | Metabolomics Quenching method Lactobacillus bulgaricus Leakage |
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