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利用高分辨率质谱鉴定的靶向肽标记物对巴西橡胶榕( Mimosa scabrella Bentham )蜜露蜜蛋白质进行定量比较和分化。

Comparative quantification and differentiation of bracatinga (Mimosa scabrella Bentham) honeydew honey proteins using targeted peptide markers identified by high-resolution mass spectrometry.

机构信息

Department of Food Science and Technology, Federal University of Santa Catarina, Florianopolis, SC, Brazil; Institute of Nutritional Science, University of Potsdam, Nuthetal OT Bergholz-Rehbrücke, Germany.

Department of Food Science and Technology, Federal University of Santa Catarina, Florianopolis, SC, Brazil.

出版信息

Food Res Int. 2021 Mar;141:109991. doi: 10.1016/j.foodres.2020.109991. Epub 2020 Dec 16.

Abstract

Honey traceability is an important topic, especially for honeydew honeys, due to the increased incidence of adulteration. This study aimed to establish specific markers to quantify proteins in honey. A proteomics strategy to identify marker peptides from bracatinga honeydew honey was therefore developed. The proteomics approach was based on initial untargeted identification of honey proteins and peptides by LC-ESI-Triple-TOF-MS/MS, which identified the major royal jelly proteins (MRJP) presence. Afterwards, the peptides were selected by the in silico digestion. The marker peptides were quantified by the developed targeted LC-QqQ-MS/MS method, which provided good linearity and specificity, besides recoveries between 92 and 100% to quantify peptides from bracatinga honeydew honey. The uniqueness and high response in mass spectrometry were backed by further complementary protein analysis (SDS-PAGE). The selected marker peptides EALPHVPIFDR (MRJP 1), ILGANVK (MRJP 2), TFVTIER (MRJP 3), QNIDVVAR (MRJP 4), FINNDYNFNEVNFR (MRJP 5) and LLQPYPDWSWTK (MRJP 7), quantified by LC-QqQ-MS/MS, highlighted that the content of QNIDVVAR from MRJP 4 could be used to differentiate bracatinga honeydew honey from floral honeys (p < 0.05) as a potential marker for its authentication. Finally, principal components analysis highlighted the QNIDVVAR content as a good descriptor of the analyzed bracatinga honeydew honey samples.

摘要

蜂蜜的溯源性是一个重要的话题,特别是对于甘露蜜,因为其掺假的情况有所增加。本研究旨在建立特定的标志物来定量分析蜂蜜中的蛋白质。因此,开发了一种从巴西橡胶榕蜜中鉴定标记肽的蛋白质组学策略。该蛋白质组学方法基于 LC-ESI-Triple-TOF-MS/MS 对蜂蜜蛋白和肽进行初始非靶向鉴定,从而鉴定出主要蜂王浆蛋白(MRJP)的存在。之后,通过计算机模拟消化选择肽。通过开发的靶向 LC-QqQ-MS/MS 方法对标记肽进行定量,该方法具有良好的线性和特异性,回收率在 92%至 100%之间,可定量巴西橡胶榕蜜中的肽。通过进一步的互补蛋白质分析(SDS-PAGE),这些肽在质谱中的独特性和高响应得到了支持。选择的标记肽 EALPHVPIFDR(MRJP1)、ILGANVK(MRJP2)、TFVTIER(MRJP3)、QNIDVVAR(MRJP4)、FINNDYNFNEVNFR(MRJP5)和 LLQPYPDWSWTK(MRJP7)通过 LC-QqQ-MS/MS 定量,突出了 MRJP4 中的 QNIDVVAR 含量可用于区分巴西橡胶榕蜜和花蜜(p<0.05),作为其鉴定的潜在标志物。最后,主成分分析突出了 QNIDVVAR 含量作为分析巴西橡胶榕蜜样品的良好描述符。

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