Chair of Food Chemistry and Molecular Sensory Science , Technische Universität München , Lise-Meitner-Strasse 34 , D-85354 Freising-Weihenstephan , Germany.
Leibniz-Institute for Food Systems Biology at the Technical University of Munich , Lise-Meitner-Strasse 34 , D-85354 Freising-Weihenstephan , Germany.
J Agric Food Chem. 2019 Aug 7;67(31):8500-8509. doi: 10.1021/acs.jafc.9b03237. Epub 2019 Jul 24.
To map qualitative and quantitative metabolome alterations when is grown in an environment where l-tyrosine levels are perturbed, the recently established differential off-line LC-NMR (DOLC-NMR) approach was successfully applied in connection with an absolute metabolite quantitation using a quantitative H NMR protocol following the ERETIC 2 (Electronic REference To access In vivo Concentrations) methodology. Among the 23 influenced metabolites, amino acid degradation products like 2-(4-hydroxyphenyl)acetic acid and 2-(3,4-dihydroxyphenyl)acetic acid underwent a tremendous upregulation in the amino acid perturbed approach. Moreover, the output of secondary metabolites like andrastin A, eremofortin B, and the tetrapeptide d-Phe-l-Val-d-Val-l-Tyr was affected in the case of the presence or absence of the added aromatic amino acid. Furthermore, the isolated secondary metabolites of have been quantified for the first time in five divergent isolates by means of a validated LC-ECHO-MS/MS method. This technique is used to compensate the effect of co-extracted matrix compounds during the analysis and to utilize quasi-internal standards to quantify all metabolites of interest accurately. This screening outlined the great variety between the different fungi of the same species. The metabolite spectra of wild-type fungi included more toxic intermediates compared to a selected fungi used as a starter culture for blue-mold cheese production. In addition, these secondary metabolites were quantified in commercially available white- and blue-mold cheese samples. The main differences between the analyte profiles of white and blue cheeses were linked to the impact of the used starter culture. Specific metabolites detected from like andrastin A and B or roquefortine C could not be detected in white cheese. Among the blue cheese samples, different metabolite pattern could be observed regarding various starter cultures.
为了绘制 在酪氨酸水平受到干扰的环境中生长时的定性和定量代谢组学变化，最近建立的差分离线 LC-NMR（DOLC-NMR）方法与使用电子参考以获取体内浓度（ERE-TIC 2）方法的定量 H NMR 方案进行绝对代谢物定量相结合，成功应用。在受影响的 23 种代谢物中，氨基酸降解产物，如 2-（4-羟苯基）乙酸和 2-（3,4-二羟苯基）乙酸，在氨基酸受扰方法中经历了巨大的上调。此外，在添加或不添加芳香族氨基酸的情况下，次级代谢产物如 andrastin A、eremofortin B 和四肽 d-Phe-l-Val-d-Val-l-Tyr 的产量也受到了影响。此外，首次通过验证的 LC-ECHO-MS/MS 方法对五种不同 分离株的次级代谢产物进行了定量。该技术用于补偿分析过程中共同提取基质化合物的影响，并利用准内标准确地定量所有感兴趣的代谢物。该筛选概述了同一物种不同真菌之间的巨大差异。与作为蓝纹奶酪生产起始培养物选择的真菌相比，野生型真菌的代谢物谱包含更多的有毒中间体。此外，这些次级代谢产物在市售的白霉和蓝霉奶酪样品中进行了定量。白奶酪和蓝奶酪分析物图谱之间的主要差异与所使用的起始培养物的影响有关。从 中检测到的特定代谢物，如 andrastin A 和 B 或 roquefortine C，在白奶酪中无法检测到。在蓝奶酪样品中，不同的代谢物模式可以观察到不同的 起始培养物。
