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CLC number: Q819

On-line Access: 2014-04-06

Received: 2013-05-24

Revision Accepted: 2013-11-18

Crosschecked: 2014-03-18

Cited: 5

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Citations:  Bibtex RefMan EndNote GB/T7714

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Journal of Zhejiang University SCIENCE B 2014 Vol.15 No.4 P.333-342

http://doi.org/10.1631/jzus.B1300149


Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus *


Author(s):  Ming-ming Chen1, Ai-li Li1, Mao-cheng Sun2, Zhen Feng1, Xiang-chen Meng1, Ying Wang1

Affiliation(s):  1. MOE Key Laboratory of Dairy Science, College of Food Science, Northeast Agricultural University, Harbin 150030, China; more

Corresponding email(s):   aili-mail@163.com

Key Words:  Metabolomics, Quenching method, Lactobacillus bulgaricus , Leakage


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 B, 2014, 15(4): 333-342.

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author="Ming-ming Chen, Ai-li Li, Mao-cheng Sun, Zhen Feng, Xiang-chen Meng, Ying Wang",
journal="Journal of Zhejiang University Science B",
volume="15",
number="4",
pages="333-342",
year="2014",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1300149"
}

%0 Journal Article
%T Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus
%A Ming-ming Chen
%A Ai-li Li
%A Mao-cheng Sun
%A Zhen Feng
%A Xiang-chen Meng
%A Ying Wang
%J Journal of Zhejiang University SCIENCE B
%V 15
%N 4
%P 333-342
%@ 1673-1581
%D 2014
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1300149

TY - JOUR
T1 - Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus
A1 - Ming-ming Chen
A1 - Ai-li Li
A1 - Mao-cheng Sun
A1 - Zhen Feng
A1 - Xiang-chen Meng
A1 - Ying Wang
J0 - Journal of Zhejiang University Science B
VL - 15
IS - 4
SP - 333
EP - 342
%@ 1673-1581
Y1 - 2014
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1300149


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.

保加利亚乳杆菌代谢组学中淬灭方法的优化研究

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。
代谢组学;淬灭处理;保加利亚乳杆菌;泄露

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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