Sathe Shridhar K, Venkatachalam Mahesh, Sharma Girdhari M, Kshirsagar Harshal H, Teuber Suzanne S, Roux Kenneth H
Department of Nutrition, Food and Exercise Sciences, Florida State University, Tallahassee, Florida 32306-1493, USA.
J Agric Food Chem. 2009 Sep 9;57(17):7846-56. doi: 10.1021/jf9016338.
The solubility of almond, Brazil nut, cashew nut, hazelnut, macadamia, pecan, pine nut, pistachio, walnut, and peanut proteins in several aqueous solvents was qualitatively and quantitatively assessed. In addition, the effects of extraction time and ionic strength on protein solubility were also investigated. Electrophoresis and protein determination (Lowry, Bradford, and micro-Kjeldahl) methods were used for qualitative and quantitative assessment of proteins, respectively. Depending on the seed, buffer type and ionic strength significantly affected protein solubility. The results suggest that buffered sodium borate (BSB; 0.1 M H(3)BO(3), 0.025 M Na(2)B(4)O(7), 0.075 M NaCl, pH 8.45) optimally solubilizes nut seed proteins. Qualitative differences in seed protein electrophoretic profiles were revealed. For a specific seed type, these differences were dependent on the solvent(s) used to solubilize the seed proteins. SDS-PAGE results suggest the polypeptide molecular mass range for the tree nut seed proteins to be 3-100 kDa. The results of native IEF suggested that the proteins were mainly acidic, with a pI range from >4.5 to <7.0. Western immunoblotting experiments indicated that rabbit polyclonal antibodies recognized substantially the same polypeptides as those recognized by the corresponding pooled patient sera IgE.
对杏仁、巴西坚果、腰果、榛子、澳洲坚果、山核桃、松子、开心果、核桃和花生蛋白在几种水性溶剂中的溶解度进行了定性和定量评估。此外，还研究了提取时间和离子强度对蛋白溶解度的影响。分别采用电泳法和蛋白质测定法（洛瑞法、布拉德福德法和微量凯氏定氮法）对蛋白质进行定性和定量评估。根据种子种类的不同，缓冲液类型和离子强度对蛋白质溶解度有显著影响。结果表明，硼酸钠缓冲液（BSB；0.1M H₃BO₃、0.025M Na₂B₄O₇、0.075M NaCl，pH 8.45）能使坚果种子蛋白达到最佳溶解度。揭示了种子蛋白电泳图谱的定性差异。对于特定的种子类型，这些差异取决于用于溶解种子蛋白的溶剂。SDS-PAGE结果表明，坚果种子蛋白的多肽分子量范围为3-100kDa。天然IEF结果表明，这些蛋白质主要呈酸性，pI范围为>4.5至<7.0。Western免疫印迹实验表明，兔多克隆抗体识别的多肽与相应的混合患者血清IgE识别的多肽基本相同。
