Gas chromatographic metabolic profiling: A sensitive tool for functional microbial ecology
Gas chromatographic metabolic profiling: A sensitive tool for functional microbial ecology
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DOI：
10.1016/j.mimet.2008.07.029
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发表时间：
2008-12-01
影响因子：
2.2
通讯作者：
Nunan, Naoise
中科院分区：
文献类型：
--
作者：
Coucheney, Elsa;Daniell, Tim J.;Nunan, Naoise
微生物代谢组学包括对细胞释放的（“外代谢组”）或存在于（“内代谢组”）的代谢物进行非靶向分析，主要用于研究特定微生物的代谢。代谢组还代表了微生物活动的图景，我们认为外代谢组也可能包含用于研究微生物相互作用网络的相关信息。气相色谱与质谱联用是代谢组学研究中最常用的技术。它可以检测多种代谢物，但需要在检测前对化合物进行衍生化。这种类型的非靶向分析可能会给不同代谢物的检测和定量带来偏差，特别是在提取和衍生化步骤中。因此，本研究的目的是量化变异性的来源，并测试 GC 代谢分析方法对温度变化等微小环境变化的敏感性。将代谢曲线的温度敏感性与使用Biolog (R) 微孔板获得的分解代谢曲线进行比较。通过在 20°C 下孵育从土壤中分离的细菌菌株与果糖，并通过重复方案的每个步骤（孵育、提取和衍生化），将分析变异性与生物变异性进行比较。对于内代谢组和外代谢组，超过 70% 的总变异性是生物起源的，主成分分析沿第一排序轴清楚地分离了菌株。内代谢组仅在物种水平上区分细菌菌株，而外代谢组在物种和群体水平上分离是明显的。温度对代谢物产生或细菌菌株具有显着但不同的影响，而它们的分解代谢特征相对不受影响。外代谢组比内代谢组对温度变化更敏感，这表明该库可能对环境功能生态学的研究感兴趣。 (C) 2008 Elsevier B.V. 保留所有权利。
Microbial metabolomics, which consists of a non-targeted analysis of the metabolites released from ('exometabolome') or existing in ('endometabolome') a cell has mostly been used to study the metabolism of particular microbes. Metabolomes also represent a picture of microbial activity and we suggest that the exometabolome may also contain pertinent information for studying microbial interaction networks. Gas chromatography coupled to mass spectrometry is the most commonly used technique in metabolomics studies. It allows a wide range of metabolites to be detected but requires the derivatisation of compounds prior to detection. This type of non-targeted analysis can introduce biases to the detection and quantification of the different metabolites, particularly at the extraction and derivatisation steps. The aims of this study, therefore, were to quantify the sources of variability and to test the sensitivity of the GC metabolic profiling approach to small environmental changes such as shifts in temperature. The temperature sensitivity of metabolic profiles was compared with that of catabolic profiles obtained using Biolog (R) microplates. Analytical variability was compared with biological variability by incubating bacterial strains isolated from soil with fructose at 20 degrees C and by replicating each step of the protocol (incubation, extraction and derivatisation). For both the endo- and the exometabolome, more than 70% of the total variability was of biological origin and principal components analysis clearly separated the strains along the first ordination axis. The endometabolome distinguished bacterial strains at the species level only, whereas separation was evident at the species and group level with the exometabolome. Temperature had a significant but differential effect on the metabolite production or the bacterial strains whilst their catabolic profiles remained relatively unaffected. The exometabolome was more sensitive to temperature shifts than the enclometabolome, suggesting that this pool may be of interest for studies in environmental functional ecology. (C) 2008 Elsevier B.V. All rights reserved.