Ferreira António César Silva, Monforte Ana Rita, Teixeira Carla Silva, Martins Rosa, Fairbairn Samantha, Bauer Florian F
Escola Superior de Biotecnologia, Universidade Católica Portuguesa , Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
J Agric Food Chem. 2014 Jul 16;62(28):6784-93. doi: 10.1021/jf502082z. Epub 2014 Jul 8.
This work describes the utility and efficiency of a metabolic profiling pipeline that relies on an unsupervised and untargeted approach applied to a HS-SPME/GC-MS data. This noninvasive and high throughput methodology enables "real time" monitoring of the metabolic changes inherent to the biochemical dynamics of a perturbed complex biological system and the extraction of molecular candidates that are latter validated on its biochemical context. To evaluate the efficiency of the pipeline five different fermentations, carried on a synthetic media and whose perturbation was the nitrogen source, were performed in 5 and 500 mL. The smaller volume fermentations were monitored online by HS-SPME/GC-MS, allowing to obtain metabolic profiles and molecular candidates time expression. Nontarget analysis was applied using MS data in two ways: (i) one dimension (1D), where the total ion chromatogram per sample was used, (ii) two dimensions (2D), where the integrity time vs m/z per sample was used. Results indicate that the 2D procedure captured the relevant information more efficiently than the 1D. It was also seen that although there were differences in the fermentation performance in different scales, the metabolic pathways responsible for production of metabolites that impact the quality of the volatile fraction was unaffected, so the proposed pipeline is suitable for the study of different fermentation systems that can undergo subsequent sensory validation on a larger scale.
这项工作描述了一种代谢谱分析流程的实用性和效率，该流程依赖于一种应用于HS-SPME/GC-MS数据的无监督且非靶向方法。这种非侵入性的高通量方法能够对受干扰的复杂生物系统生化动力学固有的代谢变化进行“实时”监测，并提取分子候选物，这些候选物随后在其生化背景下得到验证。为了评估该流程的效率，在合成培养基上进行了五种不同的发酵，其干扰因素为氮源，发酵体积分别为5 mL和500 mL。较小体积的发酵通过HS-SPME/GC-MS进行在线监测，从而获得代谢谱和分子候选物的时间表达。非靶向分析使用质谱数据的方式有两种：（i）一维（1D），即使用每个样品的总离子色谱图；（ii）二维（2D），即使用每个样品的保留时间与质荷比。结果表明，二维方法比一维方法更有效地捕捉到了相关信息。还可以看出，尽管不同规模的发酵性能存在差异，但影响挥发性成分质量的代谢产物产生所涉及的代谢途径并未受到影响，因此所提出的流程适用于不同发酵系统的研究，这些发酵系统随后可在更大规模上进行感官验证。
