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Simply and effectively preparing high‐purity phosvitin using polyethylene glycol and anion‐exchange chromatography
Author(s) -
Zhang Xiaowei,
Qiu Ning,
Geng Fang,
Ma Meihu
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100601
Subject(s) - phosvitin , polyethylene glycol , chromatography , chemistry , peg ratio , ion exchange , yield (engineering) , metal ions in aqueous solution , metal , ion , materials science , organic chemistry , protein kinase a , finance , economics , metallurgy , enzyme
An attempt was made to develop a new protocol for preparing phosvitin that could be easy scaled up using polyethylene glycol (PEG6000). Influence of PEG6000 concentration and pH of sample solution on phosvitin isolation was investigated. Phosvitin of high purity (99%) was obtained in good yield (47%) with the optimal condition of pH 4.0 and 3% PEG6000 precipitation. In addition, through evaluating different anion‐exchange chromatography methods, the DEAE procedure at pH 7.5 was finally selected as the best procedure to obtain metal‐free phosvitin that lost the least protein. Furthermore, it is observed that the purity and characterizations of prepared phosvitin were similar to those of the standard phosvitin from Sigma, and the random coil of phosvitin converted into more compact structure after removing metal ion. In conclusion, the developed method was simple and suitable for scaled up preparation of phosvitin.