New monitoring approach for metabolic dynamics in microbial ecosystems using stable-isotope-labeling technologies
New monitoring approach for metabolic dynamics in microbial ecosystems using stable-isotope-labeling technologies
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DOI：
10.1016/j.jbiosc.2010.01.004
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发表时间：
2010-07-01
影响因子：
2.8
通讯作者：
Kikuchi, Jun
中科院分区：
文献类型：
--
作者：
Date, Yasuhiro;Nakanishi, Yumiko;Kikuchi, Jun
我们已经开发了一种新的方法来监测代谢动力学的微生物生态系统中使用的DNA指纹图谱和代谢组学分析的基础上稳定同位素标记技术相结合。稳定同位素探测DNA（DNA-SIP）以前已被用于评价微生物群落中的交叉摄食。为了开发和验证我们的监测方法，使用稳定同位素标记的葡萄糖作为唯一碳源分析粪便微生物群。为了将代谢信息与微生物变异性联系起来，我们基于核磁共振（NMR）谱和变性梯度凝胶电泳（DGGE）指纹进行了代谢-微生物相关性分析，成功鉴定了葡萄糖利用菌及其相关胞外代谢产物。此外，我们的方法揭示了关于碳通量的信息，其中葡萄糖利用细菌分泌的细胞外代谢物的“第一”波被并入底物利用细菌的“次级”组，并且该“次级”组进一步产生它们自己的次级代谢底物。因此，这种方法是监测微生物生态系统中代谢动力学的有力工具，并允许跟踪微生物群落内的碳通量。(C)2010年，生物技术学会，日本。All rights reserved.
We have developed a new approach for monitoring the metabolic dynamics in microbial ecosystems using a combination of DNA fingerprinting and metabolome analysis based on stable-isotope-labeling technologies. Stable-isotope probing of DNA (DNA-SIP) has been used previously for the evaluation of cross-feeding in microbial communities. For the development and validation of our monitoring approach, fecal microbiota were analyzed with stable-isotope-labeled glucose used as the sole carbon source. In order to link the metabolic information and the microbial variability, we performed metabolic-microbial correlation analysis based on nuclear magnetic resonance (NMR) profiles and denaturing gradient gel electrophoresis (DGGE) fingerprints, which successfully identified the glucose-utilizing bacteria and their related extracellular metabolites. Moreover, our approach revealed information regarding the carbon flux, in that the "first" wave of extracellular metabolites secreted by the glucose-utilizing bacteria were incorporated into the "secondary" group of substrate-utilizing bacteria, and that this "secondary" group further produced their own secondary metabolized substrates. Thus, this approach is a powerful tool for monitoring the metabolic dynamics in microbial ecosystems and allows for the tracking of the carbon flux within a microbial community. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.