Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
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DOI：
10.1371/journal.pone.0004893
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Kikuchi J
中科院分区：
文献类型：
--
作者：
Fukuda S;Nakanishi Y;Chikayama E;Ohno H;Hino T;Kikuchi J
生态系统中的环境过程会因微生物的多种代谢反应而动态改变，这些反应包括细胞内感应和泵作用、生存竞争以及营养物质的供给或竞争。值得注意的是，肠道细菌通过多种动态生化反应维持哺乳动物体内的稳态平衡，从未消化的食物中产生多种代谢物，这些代谢物以时间依赖的方式对哺乳动物细胞产生多种影响。我们已经建立了一种实时分析细菌代谢动态的方法，并将其与统计核磁共振程序相结合。
我们开发了一种名为实时代谢分型（RT - MT）的新方法，该方法在核磁共振管中细菌生长期间进行连续的¹H - NMR谱分析和二维（2D）¹H，¹³C - HSQC（异核单量子相干）谱分析。通过Z - 分数分析、主成分分析和统计全相关光谱（TOCSY）的时间序列等统计方法对谱图进行评估。此外，利用稳定同位素标记技术进行2D ¹H，¹³C - HSQC分析，我们根据时间依赖的2D动力学谱图观察到特定生化反应的代谢动力学。利用这些方法，我们阐明了一种胃肠道细菌——溶纤维丁酸弧菌对亚麻酸的氢化途径。根据¹³C标记的RT - MT实验结果，我们确定反11，顺13共轭亚油酸是溶纤维丁酸弧菌氢化亚麻酸的中间产物。此外，我们还表明多不饱和脂肪酸的生物氢化是一种抵御其毒性作用的防御机制。
RT - MT对于表征有益细菌很有用，这些有益细菌通过产生生物活性化合物显示出用作益生菌的潜力。
Environmental processes in ecosystems are dynamically altered by several metabolic responses in microorganisms, including intracellular sensing and pumping, battle for survival, and supply of or competition for nutrients. Notably, intestinal bacteria maintain homeostatic balance in mammals via multiple dynamic biochemical reactions to produce several metabolites from undigested food, and those metabolites exert various effects on mammalian cells in a time-dependent manner. We have established a method for the analysis of bacterial metabolic dynamics in real time and used it in combination with statistical NMR procedures. We developed a novel method called real-time metabolotyping (RT-MT), which performs sequential 1H-NMR profiling and two-dimensional (2D) 1H, 13C-HSQC (heteronuclear single quantum coherence) profiling during bacterial growth in an NMR tube. The profiles were evaluated with such statistical methods as Z-score analysis, principal components analysis, and time series of statistical TOtal Correlation SpectroScopY (TOCSY). In addition, using 2D 1H, 13C-HSQC with the stable isotope labeling technique, we observed the metabolic kinetics of specific biochemical reactions based on time-dependent 2D kinetic profiles. Using these methods, we clarified the pathway for linolenic acid hydrogenation by a gastrointestinal bacterium, Butyrivibrio fibrisolvens. We identified trans11, cis13 conjugated linoleic acid as the intermediate of linolenic acid hydrogenation by B. fibrisolvens, based on the results of 13C-labeling RT-MT experiments. In addition, we showed that the biohydrogenation of polyunsaturated fatty acids serves as a defense mechanism against their toxic effects. RT-MT is useful for the characterization of beneficial bacterium that shows potential for use as probiotic by producing bioactive compounds.
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影响因子：
2.8
作者：
Diez-Gonzalez, F;Bond, DR;Russell, JB
通讯作者：
Russell, JB
影响因子：
4
作者：
Fukuda, S;Suzuki, Y;Hino, T
通讯作者：
Hino, T
影响因子：
56.9
作者：
Gill, Steven R.;Pop, Mihai;Nelson, Karen E.
通讯作者：
Nelson, Karen E.
DOI：
10.1152/ajpgi.90227.2008
发表时间：
2008-11-01
影响因子：
4.5
作者：
Ewaschuk, Julia B.;Diaz, Hugo;Madsen, Karen L.
通讯作者：
Madsen, Karen L.
DOI：
10.1080/03601230701795262
发表时间：
2008-02-01
影响因子：
2
作者：
Chalova, V. I.;Lingbeck, J. M.;Ricke, S. C.
通讯作者：
Ricke, S. C.