Proteomics and Metabolomics Laboratory, Crop Research Institute, Drnovska 507/73, Prague, CZ-16106, Czechia.
Proteomics and Metabolomics Laboratory, Crop Research Institute, Drnovska 507/73, Prague, CZ-16106, Czechia.
J Proteomics. 2021 May 15;239:104157. doi: 10.1016/j.jprot.2021.104157. Epub 2021 Feb 22.
Honey adulteration is a common practice that deceives consumers and devalues the unique curative and food properties of honey. For marketing, each honey must satisfy an internationally valid Codex standard. One of the quality parameters is diastase/amylase activity, which, if lowered, may be compensated for by the addition of foreign amylases. However, the estimation of enzyme activity does not enable identification of artificially added amylases. 45 honey samples were analyzed using label-free nanoLC-MS/MS proteomics. Four honeys were found to contain the foreign amylases from Aspergillus niger, Bacillus amyloliquefaciens and/or Bacillus licheniformis. This result was confirmed via proof of specificity at multiple levels. Furthermore, we identified a series of plant-related protein groups. Despite plant-related proteins constituting a significant portion of honey proteins, they were minor components compared to the major honey bee-derived proteins. Bioinformatic analysis also provided evidence for aphid and catalase proteins in honey, but the limited specificity of the MS/MS identified peptides must be considered. Overall, we demonstrate a proteomics approach employing LC-MS/MS that is useful for proving adulteration and assessing honey quality. As an resource useful for reference, we provide curated sequence databases. In addition, we provide many markers that are naturally found in honey for future studies. SIGNIFICANCE: Honey is unique natural product used since ancient times as a food and natural medicine. Humans strive to understand honey components because they can characterize different types of honey and be used for authentication and origin assessment. One of the important honey components are proteins. The proteins present in honey can naturally occur in honey, but some of them can be used to mask deficiencies in some honey quality properties. Diastases/amylases are such proteins, and their activity, a measure of honey freshness, can decrease in time or due to processing. To our knowledge, we for the first time specifically identify foreign amylases in honey. However, this study provided new information on other non-honey bee proteins in honey. Thus, this study is also of importance due to its identification of plant and aphid proteins and catalase-related proteins. This study provides a clue explaining the controversial presence of catalase in honey, since catalases can be identified and their origin determined via proteomics.
蜂蜜掺假是一种常见的做法，它欺骗了消费者，降低了蜂蜜独特的治疗和食用属性。为了营销，每种蜂蜜都必须满足国际有效的食品法典标准。质量参数之一是酶活性（如淀粉酶），如果降低，可以通过添加外源淀粉酶来补偿。然而，酶活性的估计并不能识别人工添加的淀粉酶。使用无标签的纳升 LC-MS/MS 蛋白质组学分析了 45 个蜂蜜样本。发现四种蜂蜜含有黑曲霉、解淀粉芽孢杆菌和/或地衣芽孢杆菌的外源淀粉酶。通过在多个层面证明特异性，证实了这一结果。此外，我们还鉴定了一系列与植物相关的蛋白质组。尽管与植物相关的蛋白质构成了蜂蜜蛋白质的重要部分，但与主要的蜜蜂衍生蛋白质相比，它们是次要成分。生物信息学分析还为蜂蜜中的蚜虫和过氧化氢酶蛋白提供了证据，但必须考虑到 MS/MS 鉴定肽的有限特异性。总的来说，我们展示了一种使用 LC-MS/MS 的蛋白质组学方法，该方法可用于证明掺假和评估蜂蜜质量。作为参考资源，我们提供了经过整理的序列数据库。此外，我们还提供了许多在未来研究中可用于鉴定蜂蜜的天然标志物。意义：蜂蜜是一种独特的天然产物，自古以来一直被用作食品和天然药物。人类努力了解蜂蜜的成分，因为它们可以表征不同类型的蜂蜜，并用于认证和原产地评估。蛋白质是蜂蜜的重要成分之一。存在于蜂蜜中的蛋白质可能天然存在于蜂蜜中，但其中一些蛋白质可用于掩盖某些蜂蜜质量特性的缺陷。淀粉酶就是这种蛋白质，它是衡量蜂蜜新鲜度的指标，其活性会随时间或因加工而降低。据我们所知，我们首次专门鉴定了蜂蜜中的外源淀粉酶。然而，这项研究还提供了有关蜂蜜中其他非蜜蜂蛋白的新信息。因此，由于其对植物和蚜虫蛋白以及与过氧化氢酶相关的蛋白质的鉴定，该研究也具有重要意义。这项研究提供了一个线索，可以解释过氧化氢酶在蜂蜜中存在的争议，因为通过蛋白质组学可以识别和确定其来源。
