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将 Pedobacter lusitanus NL19 挥发性外代谢组与生长培养基成分相关联:使用全面二维气相色谱法结合飞行时间质谱法,我们可以学到什么?

Linking Pedobacter lusitanus NL19 volatile exometabolome with growth medium composition: what can we learn using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry?

机构信息

Department of Biology & CESAM, University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal.

Department of Chemistry & LAQV-REQUIMTE, University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal.

出版信息

Anal Bioanal Chem. 2023 May;415(13):2613-2627. doi: 10.1007/s00216-022-04505-6. Epub 2023 Jan 11.

DOI:10.1007/s00216-022-04505-6
PMID:36631573
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10149447/
Abstract

Microbial metabolomics allows understanding and to comprehensively analyse metabolites, and their related cellular and metabolic processes, that are produced and released to the extracellular environment under specific conditions. In that regard, the main objective of this research is to understand the impact of culture media changes in the metabolic profile of Pedobacter lusitanus NL19 (NL19) and Pedobacter himalayensis MTCC 6384 (MTCC6384) and respective influence on the production of biotechnologically relevant compounds. Solid-phase microextraction combined with comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry with time-of-flight analyser (GC × GC-ToFMS) was applied to comprehensively study the metabolites produced by NL19 and MTCC6384 both in tryptic soy broth 100% (TSB100) and tryptic soy broth with 25% casein peptone (PC25). A total of 320 metabolites were putatively identified, which belong to different chemical families: alcohols, aldehydes, esters, ethers, hydrocarbons, ketones, nitrogen compounds, sulphur compounds, monoterpenes, and sesquiterpenes. Metabolites that were statistically different from the control (sterile medium) were selected allowing for the construction of the metabolic profile of both strains. A set of 80 metabolites was tentatively associated to the metabolic pathways such as the metabolism of fatty acids, branched-chain aminoacids, phenylalanine, methionine, aromatic compounds, and monoterpene and sesquiterpene biosynthesis. This study allowed to better understand how slight changes of the culture media and thus the composition of nutrients impair the metabolic profile of bacteria, which may be further explored for metabolomics pipeline construction or biotechnological applications.

摘要

微生物代谢组学可以帮助理解和全面分析代谢物,以及在特定条件下产生并释放到细胞外环境中的与其相关的细胞和代谢过程。在这方面,本研究的主要目的是了解培养介质变化对 Pedobacter lusitanus NL19 (NL19) 和 Pedobacter himalayensis MTCC 6384 (MTCC6384) 代谢谱的影响,以及它们对生物技术相关化合物生产的各自影响。固相微萃取与全二维气相色谱-飞行时间质谱联用(GC×GC-ToFMS)相结合,用于全面研究 NL19 和 MTCC6384 在胰蛋白胨大豆肉汤 100%(TSB100)和含 25%酪蛋白胨的胰蛋白胨大豆肉汤(PC25)中产生的代谢物。共鉴定出 320 种假定代谢物,它们属于不同的化学家族:醇、醛、酯、醚、烃、酮、含氮化合物、含硫化合物、单萜和倍半萜。从对照(无菌培养基)中选择具有统计学差异的代谢物,从而构建了两种菌株的代谢谱。一组 80 种代谢物被初步与脂肪酸、支链氨基酸、苯丙氨酸、蛋氨酸、芳香族化合物以及单萜和倍半萜生物合成等代谢途径相关联。本研究有助于更好地了解培养介质的微小变化以及营养成分的组成如何影响细菌的代谢谱,这可以进一步探索用于代谢组学管道构建或生物技术应用的代谢谱。

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