Department of Life Science and Medical Bioscience, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan.
J Biosci Bioeng. 2010 Jul;110(1):87-93. doi: 10.1016/j.jbiosc.2010.01.004. Epub 2010 Jan 27.
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.
我们开发了一种新的方法，用于监测微生物生态系统中的代谢动态，该方法结合了 DNA 指纹图谱和基于稳定同位素标记技术的代谢组分析。以前曾使用稳定同位素探测 DNA（DNA-SIP）来评估微生物群落中的交叉喂养。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来分析粪便微生物群。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）图谱和变性梯度凝胶电泳（DGGE）指纹图谱进行了代谢微生物相关性分析，该分析成功地鉴定了葡萄糖利用细菌及其相关的细胞外代谢物。此外，我们的方法还揭示了有关碳通量的信息，即葡萄糖利用细菌分泌的“第一”波细胞外代谢物被纳入“次级”组的底物利用细菌中，并且该“次级”组进一步产生了自己的次级代谢物底物。因此，该方法是监测微生物生态系统代谢动态的有力工具，并允许跟踪微生物群落内的碳通量。
