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Purification of Hen’s Egg Ovomucoid by Three-Step Precipitation

WANG Shuai1, WU Zi-jian1,*, WANG Su-ying1, LIU Jian-fu1, ZHANG Wei1, TIE Hang1, RONG Qiang2   

  1. 1. Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science, Tianjin University of Commerce,
    Tianjin 300134, China; 2. Cuisine Department, Tianjin Youth Professional Technology Institute, Tianjin 300191, China
  • Online:2014-12-25 Published:2014-12-29
  • Contact: WU Zi-jian

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Abstract:

Hen’s egg ovomucoid (HOVM) is the dominant allergenic protein from chicken egg white, and obtaining highpurity
HOVM is the prerequisite for understanding its structure and characteristics and reducing its allergenicity. This study
presented a three-step precipitation procedure (precipitation with 10% trichloroacetic acid (TCA) followed by two-step
fractional precipitation with ammonium sulfate) for purifying HOVM. The purification efficiency was evaluated by SDSPAGE
and HPLC. It was found that after most of the protein impurities in the 2-fold diluted egg white were removed by
adding 10% TCA, the centrifugal supernatant was added with ammonium sulfate to saturation (50%) for further removal
of non-HOVM proteins by salting-out. Finally, additional ammonium sulfate was added to the supernatant fraction until
saturation (80%). After centrifugation, the precipitation fraction was dialyzed and freeze-dried to obtain high-purity HOVM
product. HPLC analysis (at 220 nm) showed that the purity of the HOVM was 99.202%, with a yield of 27.05%.

Key words: hen’s egg ovomucoid, three-step precipitation, salting-out

CLC Number: