Untargeted metabolomics via high-resolution mass spectrometry (HRMS) can reveal more than 100,000 molecular features in a single sample, many of which may represent unidentified metabolites, posing significant challenges to data analysis. We here introduce Metaboseek, an open-source analysis platform designed for untargeted comparative metabolomics and demonstrate its utility by uncovering biosynthetic functions of a conserved fat metabolism pathway, α-oxidation, using C. elegans as a model. Metaboseek integrates modules for molecular feature detection, statistics, molecular formula prediction, and fragmentation (MS/MS) analysis, which uncovered more than 200 previously uncharacterized α-oxidation-dependent metabolites in an untargeted comparison of wildtype and α-oxidation-defective hacl-1 mutants. The identified metabolites support the predicted enzymatic function of HACL-1 and revealed that α-oxidation participates in metabolism of endogenous β-methyl-branched fatty acids and food-derived cyclopropane lipids. Our results showcase compound discovery and feature annotation at scale via untargeted comparative metabolomics applied to a conserved primary metabolic pathway and suggest a model for the metabolism of cyclopropane lipids.

中文翻译：

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使用 Metaboseek 的比较代谢组学揭示了秀丽隐杆线虫中保守的脂肪代谢途径的功能

通过高分辨率质谱 (HRMS) 进行的非靶向代谢组学可以揭示单个样品中 100,000 多个分子特征，其中许多可能代表未识别的代谢物，对数据分析构成重大挑战。我们在此介绍 Metaboseek，这是一种为非靶向比较代谢组学设计的开源分析平台，并通过使用秀丽隐杆线虫作为模型揭示保守脂肪代谢途径 α-氧化的生物合成功能来证明其效用。Metaboseek 集成了用于分子特征检测、统计、分子式预测和碎片 (MS/MS) 分析的模块，在野生型和 α-氧化缺陷型hacl-的非靶向比较中发现了 200 多种以前未表征的 α-氧化依赖性代谢物1突变体。鉴定出的代谢物支持 HACL-1 的预测酶功能，并表明 α-氧化参与内源性 β-甲基支链脂肪酸和食物衍生的环丙烷脂质的代谢。我们的结果展示了通过应用于保守的主要代谢途径的非靶向比较代谢组学进行大规模化合物发现和特征注释，并提出了环丙烷脂质代谢模型。