Nciri Nader, Cho Namjun, El Mhamdi Faiçal, Ben Ismail Hanen, Ben Mansour Abderraouf, Sassi Fayçal Haj, Ben Aissa-Fennira Fatma
1 Intestinal Immunophysiology-Research Unit (02/RU/09-02), Faculty of Medicine of Tunis, University of Tunis El Manar, Bab Saâdoun, Tunisia .
2 Department of Animal Resources, Fisheries, and Food Technology, National Institute of Agronomy of Tunisia, El Mahrajène, Tunisia .
J Med Food. 2015 Sep;18(9):1049-64. doi: 10.1089/jmf.2014.0120. Epub 2015 Mar 16.
This research aimed at assessing the content and the functional properties of phytohemagglutinin (PHA) in different varieties of beans widely consumed in Tunisia through soaking, cooking, autoclaving, germination, and their combinations. This study was carried out on three varieties of white beans grown in different localities of Tunisia, namely Twila, Coco, and Beldia, as well as on imported and local canned beans. All bean samples underwent biochemical and immunological evaluation by employing several techniques such as indirect competitive enzyme-linked immunosorbent assay (ELISA), hemagglutinating assay, Ouchterlony double immunodiffusion, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Biochemical and immunological analyses indicated that raw dry beans contained a considerable amount of proteins and PHAs. ELISA demonstrated that soaking, either in plain water or in alkaline solution, caused an increase in the concentration of PHA. A slight increase of PHA was produced equally by germination during 4 days in all bean varieties. Cooking or autoclaving of presoaked beans resulted in a complete disappearance of PHA. ELISA test also proved that both imported and local canned beans contained fingerprints of PHA. Hemagglutination assays showed that not only cooked and autoclaved presoaked beans lacked the ability to agglutinate red blood cells but also autoclaved unsoaked beans did. In agar gel immunodiffusion using rabbit anti-PHA serum, raw, soaked, cooked unsoaked, and sprouted beans gave precipitin arc reactions, indicating that PHA existed in immunoreactive form in the tested seeds. SDS-PAGE electrophoretograms showed protein isolates of Twila and Beldia beans to have different profiles through soaking, cooking, and autoclaving processes. This work revealed that the combination of soaking and cooking/autoclaving was the best way in reducing PHA content and its activity in all bean varieties when compared with germination.
本研究旨在通过浸泡、烹饪、高压灭菌、发芽及其组合方式,评估突尼斯广泛食用的不同品种豆类中植物血凝素(PHA)的含量和功能特性。本研究对在突尼斯不同地区种植的三个白豆品种,即Twila、Coco和Beldia,以及进口和本地罐装豆类进行了研究。所有豆类样品均采用间接竞争酶联免疫吸附测定(ELISA)、血凝测定、双向免疫扩散和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)等多种技术进行生化和免疫学评估。生化和免疫学分析表明,生干豆含有大量蛋白质和PHA。ELISA表明,无论是用清水还是碱性溶液浸泡,PHA的浓度都会增加。在所有豆类品种中,4天的发芽过程均使PHA略有增加。预浸泡豆类的烹饪或高压灭菌会导致PHA完全消失。ELISA试验还证明,进口和本地罐装豆类均含有PHA的痕迹。血凝试验表明,不仅预浸泡后煮熟和高压灭菌的豆类缺乏凝集红细胞的能力,未浸泡直接高压灭菌的豆类也缺乏这种能力。在使用兔抗PHA血清的琼脂凝胶免疫扩散试验中,生豆、浸泡豆、未浸泡煮熟豆和发芽豆均产生沉淀弧反应,表明在受试种子中PHA以免疫反应形式存在。SDS-PAGE电泳图谱显示,Twila和Beldia豆类的蛋白质分离物在浸泡、烹饪和高压灭菌过程中具有不同的图谱。这项工作表明,与发芽相比,浸泡和烹饪/高压灭菌相结合是降低所有豆类品种中PHA含量及其活性的最佳方法。