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通过酸性毛细管电泳从不同小麦品种中提取和分离水溶性蛋白质

Extraction and separation of water-soluble proteins from different wheat species by acidic capillary electrophoresis.

作者信息

Piergiovanni Angela R

机构信息

Istituto di Genetica Vegetale-CNR, via Amendola 165/a, 70126 Bari, Italy.

出版信息

J Agric Food Chem. 2007 May 16;55(10):3850-6. doi: 10.1021/jf063628n. Epub 2007 Apr 17.

Abstract

Optimization of protein extraction and a capillary zone electrophoresis method for water-soluble protein analysis in wheat is described. The optimal separation was obtained with a 50 microm i.d. x 27 cm (20 cm to detector) uncoated capillary filled with 0.1 M phosphoric acid/beta-alanine, pH 2.5, buffer containing urea (1 M), 0.05% (w/v) hydroxypropylmethylcellulose, and 20% (v/v) acetonitrile. Separation was carried out at 15 kV and 35 degrees C for 9 min. Extract stability was also investigated within 2 h from the extraction. Good visual peak parameters and a higher sensitivity can be obtained when 30% ethanol is used as an extraction medium. The method was successfully used to analyze extracts obtained from whole and refined meals of six Triticum spp. Moreover, the described methodology could be applied to the discrimination of species with different ploidy levels and to the detection of durum wheat adulteration, as well as to screen wheat collections for enzymes involved with the quality of wheat derivatives.

摘要

本文描述了用于小麦水溶性蛋白质分析的蛋白质提取和毛细管区带电泳方法的优化。使用内径50微米×27厘米(至检测器20厘米)的未涂层毛细管,填充0.1M磷酸/β-丙氨酸、pH 2.5、含有尿素(1M)、0.05%(w/v)羟丙基甲基纤维素和20%(v/v)乙腈的缓冲液,可获得最佳分离效果。在15 kV和35℃下进行9分钟的分离。还研究了提取后2小时内提取物的稳定性。当使用30%乙醇作为提取介质时,可获得良好的视觉峰参数和更高的灵敏度。该方法成功用于分析六种小麦属全粉和精制粉提取物。此外,所描述的方法可用于区分不同倍性水平的物种、检测硬粒小麦掺假,以及筛选小麦样品中与小麦衍生物品质相关的酶。

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