Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, T6G 2P5 Alberta, Canada.
J Chromatogr A. 2012 Feb 3;1223:41-6. doi: 10.1016/j.chroma.2011.12.023. Epub 2011 Dec 13.
The objective of this study was to develop a simple method of phosvitin purification from hen egg yolk without using organic solvents. Egg yolk was diluted with equal volume of water and stirred for one hour at room temperature, followed by centrifugation to remove soluble proteins along with most of the yolk lipids in the supernatant. The granules were collected as the precipitate containing minimum amount of lipids (dry granules). The dry granules were dissolved in 0.05 M carbonate-bicarbonate buffer at pH 9.6, which yields a light yellowish solution used for anion exchange chromatography. Phosvitin fraction was collected from anion exchange chromatography as the last eluting peak with a purity of 92.6% and a yield of 35.4% of total phosvitin in the yolk or a recovery of 1.9% of total yolk dry matter, which are comparable to current methods employing organic solvents or chromatography after salt fractionation and dialysis. This method developed is simple and fast without using organic solvents.
本研究旨在开发一种从鸡蛋黄中提取卵黄高磷蛋白的简单方法,不使用有机溶剂。将蛋黄与等体积的水稀释,在室温下搅拌 1 小时,然后离心去除上清液中的可溶性蛋白质和大部分蛋黄脂质。沉淀物为含有最少脂质的颗粒(干颗粒)。将干颗粒溶解在 0.05 M 碳酸盐-碳酸氢盐缓冲液中,pH 值为 9.6,得到浅黄色溶液,用于阴离子交换层析。将卵黄高磷蛋白从阴离子交换层析中收集作为最后洗脱峰,纯度为 92.6%,收率为蛋黄中总卵黄高磷蛋白的 35.4%,或总蛋黄干物质的 1.9%,与当前使用有机溶剂或盐析和透析后层析的方法相当。该方法简单快速,不使用有机溶剂。