Institute of Clinical Chemistry and Laboratory Medicine, Section Mass Spectrometry and Proteomics, University Medical Center Hamburg-Eppendorf, Martinistraße 52, 20246 Hamburg, Germany.
Int J Mol Sci. 2021 Nov 30;22(23):12999. doi: 10.3390/ijms222312999.
Truffles of the species are known as expensive foods, mainly for their distinct aroma and taste. This high price makes them a profitable target of food fraud, e.g., the misdeclaration of cheaper truffle species as expensive ones. While many studies investigated truffles on the metabolomic level or the volatile organic compounds extruded by them, research at the proteome level as a phenotype determining basis is limited. In this study, a bottom-up proteomic approach based on LC-MS/MS measurements in data-independent acquisition mode was performed to analyze the truffle species and , and a protein atlas of the investigated species was obtained. The yielded proteomic fingerprints are unique for each of the of the five truffle species and can now be used in case of suspected food fraud. First, a comprehensive spectral library containing 9000 proteins and 50,000 peptides was generated by two-dimensional liquid chromatography coupled to tandem mass spectrometry (2D-LC-MS/MS). Then, samples of the truffle species were analyzed in data-independent acquisition (DIA) proteomics mode yielding 2715 quantified proteins present in all truffle samples. Individual species were clearly distinguishable by principal component analysis (PCA). Quantitative proteome fingerprints were generated from 2066 ANOVA significant proteins, and side-by-side comparisons of truffles were done by T-tests. A further aim of this study was the annotation of functions for the identified proteins. For and conclusive links to their superior aroma were found by enrichment of proteins responsible for sulfur-metabolic processes in comparison with other truffles. The obtained data in this study may serve as a reference library for food analysis laboratories in the future to tackle food fraud by misdeclaration of truffles. Further identified proteins with their corresponding abundance values in the different truffle species may serve as potential protein markers in the establishment of targeted analysis methods. Lastly, the obtained data may serve in the future as a basis for deciphering the biochemistry of truffles more deeply as well, when protein databases of the different truffle species will be more complete.
松露属的物种被认为是昂贵的食品，主要因其独特的香气和味道。这种高价格使它们成为食品欺诈的有利可图目标，例如，将较便宜的松露物种误报为昂贵的物种。虽然许多研究都在代谢组学水平或它们挤出的挥发性有机化合物上研究了松露，但在作为表型决定基础的蛋白质组学水平上的研究是有限的。在这项研究中，采用基于 LC-MS/MS 测量的自上而下的蛋白质组学方法，在数据非依赖性采集模式下对 和 进行分析，获得了所研究物种的蛋白质图谱。产生的蛋白质组指纹图谱对每种五种松露物种都是独特的，现在可以在怀疑食品欺诈的情况下使用。首先，通过二维液相色谱与串联质谱（2D-LC-MS/MS）生成包含 9000 种蛋白质和 50000 种肽的综合光谱库。然后，在数据非依赖性采集（DIA）蛋白质组学模式下分析松露物种的样品，得到存在于所有松露样品中的 2715 种定量蛋白质。主成分分析（PCA）清楚地区分了各个物种。从 2066 个通过 ANOVA 显著的蛋白质中生成了定量蛋白质指纹图谱，并通过 T 检验对松露进行了并排比较。本研究的另一个目的是对鉴定出的蛋白质进行功能注释。与其他松露相比，在比较硫代谢过程中负责的蛋白质的富集中，发现了 和 与其卓越香气的明确联系。本研究获得的数据将来可作为食品分析实验室的参考文库，以解决因松露误报而导致的食品欺诈问题。在不同的松露物种中，具有相应丰度值的进一步鉴定出的蛋白质可以作为建立靶向分析方法的潜在蛋白质标记。最后，当不同松露物种的蛋白质数据库更加完整时，未来获得的数据也可以作为更深入解析松露生物化学的基础。
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