Food Allergy Research and Resource Program, Department of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, Nebraska 68588, United States.
J Proteome Res. 2022 Jul 1;21(7):1694-1706. doi: 10.1021/acs.jproteome.2c00095. Epub 2022 Jun 3.
Cashews are one of the most prevalent causes of tree nut allergies. However, the cashew proteome is far from complete, which limits the quality of peptide identification in mass spectrometric analyses. In this study, bioinformatics tools were utilized to construct a customized cashew protein database and improve sequence quality for proteins of interest, based on a publicly available cashew genome database. As a result, two additional isoforms for cashew 2S albumins and five other isoforms for cashew 11S proteins were identified, along with several other potential allergens. Using the optimized protein database, the protein profiles of cashew nuts subjected to different oil-roasting conditions (139 and 166 °C for 2-10 min) were analyzed using discovery LC-MS/MS analysis. The results showed that the cashew 2S protein is most heat-stable, followed by 11S and 7S proteins, though protein isoforms might be affected differently. Preliminary target peptide selection indicated that out of the 29 potential targets, 18 peptides were derived from the newly developed database. In the evaluation of thermal processing effects on cashew proteins, several Maillard reaction adducts were also identified. The cashew protein database developed in this study allows for comprehensive analyses of cashew proteins and development of high-quality allergen detection methods.
腰果是最常见的导致坚果过敏的过敏原之一。然而,腰果蛋白组远未完全解析,这限制了质谱分析中肽鉴定的质量。在这项研究中,基于公开的腰果基因组数据库,利用生物信息学工具构建了一个定制的腰果蛋白数据库,并提高了目标蛋白的序列质量。结果鉴定出了两个腰果 2S 白蛋白的额外同型物和五个腰果 11S 蛋白的其他同型物,以及其他几种潜在的过敏原。使用优化后的蛋白数据库,对经过不同油烤条件(139 和 166°C 分别烘烤 2-10 分钟)的腰果坚果的蛋白图谱进行了发现 LC-MS/MS 分析。结果表明,腰果 2S 蛋白最稳定,其次是 11S 和 7S 蛋白,但蛋白同型物可能受到不同的影响。初步的目标肽选择表明,在 29 个潜在目标中,有 18 个肽来自新开发的数据库。在评估热加工对腰果蛋白的影响时,还鉴定出了几种美拉德反应加合物。本研究中开发的腰果蛋白数据库可用于全面分析腰果蛋白,并开发高质量的过敏原检测方法。