短短芽胞杆菌代谢物组测定中代谢终止方式的优化

陈梅春; 刘波; 朱育菁; 陈峥; 史怀; 刘国红; 潘志针

doi:10.19303/j.issn.1008-0384.2012.12.017

短短芽胞杆菌代谢物组测定中代谢终止方式的优化

doi: 10.19303/j.issn.1008-0384.2012.12.017

福建省农业科学院农业生物资源研究所,福建福州,350003

基金项目:

国家“948”项目(2011-G25)

福建省青年科技人才创新基金项目(2011J05053)

国家重点基础研究发展计划(“973”计划)前期项目(2011CB111607)

福建省农业科学院青年科技创新基金(2012DBS-1)

详细信息

作者简介:
陈梅春(1985-),女,博士,助理研究员,主要从事微生物代谢组学研究;刘波(1957-),博士,男,研究员,主要从事微生物生物技术与农业生物药物研究(E-mail:fzliubo@163.com)

中图分类号: S476.1
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- 文章访问数: 164
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- PDF下载量: 2
- 被引次数: 0
出版历程
- 收稿日期: 2012-10-24
- 刊出日期: 2012-12-18

Metabolomic Quenching for Brevibacillus brevis

Agricultural Bioresources Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou, Fujian 350003,China

摘要

摘要: 微生物代谢组学是定性和定量研究分析细胞内外所有低分子量代谢产物,在发酵控制分析、代谢物控制分析及其他组学联用进行菌种改良等方面得到了应用。胞内代谢物种类和浓度变化非常迅速,因此快速冻结代谢反应是非常重要的,所以必须有效率高和可靠的代谢终止方式,减少代谢物的流失。本研究以短短芽胞杆菌JK-2为研究对象,以LC-MS测定为评价标准,考察了5种代谢终止方式对胞内代谢物保留程度。结果表明,冷甲醇/水(含HEPES)代谢终止方式能够保留的代谢物最多,效率最高及其重现性好,在LC-MS正离子模式下,能够提取出3 000多种胞内代谢物。同时也考察了样品在正负模式下测试情况,结果表明正离子模式下能够检测到的代谢物的数目远远超过负离子模式下检测的数目。
- 短短芽胞杆菌 /
- 代谢组学 /
- 灭活 /
- 胞内代谢物 /
- 正负离子模式
Abstract: Microbial metabolomics refers to the qualitative and quantitative analysis of all low molecular weight metabolites present in and around the microbial cells.It has been widely applied in combination with other biological fields of studies for fermentation process control,metabolites analysis and microbial species upgrading.In cultivating microorganisms,the intracellular metabolites can turnover in an extremely rapid pace.Consequently,instantaneous fixation of the metabolism is essential for an accurate sampling.And,the development of a highly efficient and reliable quenching protocol is in order for most metabolomical studies.In this study,5 quenching solvent mixtures were evaluated by comparing their intracellular metabolite retention rates measured by LC-MS using the cold methanol extraction method.The results indicated that cold-methanol/water = 3/2(V/V) containing the buffer agent,HEPES,was the most efficient and reproducible quenching agent for Brevibacillus brevis cell culture.Using this method of quenching,more than 3 000 intracellular metabolites were detected in the positive ion ESI mode,which could detect more metabolites than the negative ion ESI mode.
- Brevibacillus brevis /
- metabonomics /
- quenching /
- intracellular metabolites /
- positive and negative ions ESI modes

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