The amount of cold press oil manufacture is globally rising, which in turn leads to the accumulation of deoiled plant seeds at significant quantities and consequent manufacture of plant protein products. In this study, we made an attempt to analyze the protein profile of black cumin seed protein concentrates prepared by the alkali extraction-acid precipitation technique (AE-IP). The analytical strategy relied on gel-based proteome mapping which included two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF/TOF). 14 different protein bands were identified, and in gel-trypsinolysis was carried out for the corresponding gel spots. Using the MASCOT database, current findings on 10 proteins were compared with the existing data. The highest similarity was 46 which was obtained between the highest pI black cumin protein observed here and the cyclin dependent kinase inhibitor of Arabidopsis thaliana. The molecular mass of the intact protein was determined by linear MALDI-TOF/TOF-MS as 23,711.2186 Da. The peptide constructs of this protein have been further studied in order to identify potential biological activity. Matching sequences generated bioactive peptides in silico such as IR, AL, and SL dipeptides during sequential enzymatic digestion with pepsin and trypsin. Since the majority of bioactivity investigations on black cumin seeds have been related to black cumin oil and its oil soluble components, the structure and bioactivities of black cumin proteins deserve further research.

中文翻译：

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基于二维凝胶电泳和 MALDI-TOF/TOF-MS 分析从黑孜然籽粕中鉴定新型蛋白质

冷榨油的制造量在全球范围内不断增加，这反过来又导致大量脱油植物种子的积累以及随之而来的植物蛋白产品的制造。在本研究中，我们尝试分析通过碱提取-酸沉淀技术 (AE-IP) 制备的黑孜然籽浓缩蛋白的蛋白质谱。分析策略依赖于基于凝胶的蛋白质组图谱，其中包括二维凝胶电泳，然后是基质辅助激光解吸/电离飞行时间 (MALDI-TOF/TOF)。鉴定了14条不同的蛋白条带，并对相应的凝胶点进行了凝胶胰蛋白酶分解。使用 MASCOT 数据库，将当前对 10 种蛋白质的发现与现有数据进行了比较。最高的相似性是 46，这是在此处观察到的最高 pI 黑孜然蛋白与拟南芥的细胞周期蛋白依赖性激酶抑制剂之间获得的。完整蛋白质的分子量通过线性 MALDI-TOF/TOF-MS 测定为 23,711.2186 Da。已经进一步研究了该蛋白质的肽构建体以鉴定潜在的生物活性。在用胃蛋白酶和胰蛋白酶进行连续酶消化期间，匹配序列在计算机上生成了生物活性肽，例如 IR、AL 和 SL 二肽。由于对黑孜然种子的大部分生物活性研究都与黑孜然油及其油溶性成分有关，因此黑孜然蛋白的结构和生物活性值得进一步研究。