Purification of ovalbumin from hen egg white by high-speed counter-current aqueous two-phase chromatography

Wenbo Zhi, Qiu Yun Deng, Jiang Nan Song, Fan Ouyang

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

High-speed counte-recurrent chromatography (HSCCC) is a continuous liquid-liquid partition chromatography without solid matrix, which has the significant features of high resolution and high recovery. The separation of bio-macromolecule in aqueous two-phase systems (ATPs) with HSCCC is still under research, and the establishment of high-speed counter-current aqueous two-phase chromatography (HSCCC-ATP) relies on the improvement of equipment structure and optimization of operation parameters. By using a multi-column high-speed counter-current chromatograph, the separation of protein mixture and the purification of ovalbumin from hen egg white were studied. The effects of pH and PEG concentration on the partition coefficients of proteins were tested in PEG1000-phosphate ATPs, and distinct differences among partition coefficients of proteins were found at pH 9.2 and 15.0% (W/W) PEG concentration in said system. The separation of protein mixture, consisting of cytochrome C, lysozyme and myoglobin was successfully performed in 15.0% (W/W) PEG1000-17.0% (W/W) potassium phosphate ATPs at pH 9.2 with high-speed counter-current chromatograph at rotation speed of 850r/min and flow rate of 0.8mL/min, using upper phase as stationary phase. pH and PEG concentration also had distinct effects on the partition coefficients of the major protein components in hen egg white, including ovaltransferrin, ovalbumin and lysozyme. The optimal pH value and PEG concentration for the purification of ovalbumin by HSCCC-ATP were found to be 9.2 and 16.0% (W/W) respectively. Ovalbumin was successfully purified to homogeneity from the hen egg white sample in 16.0% (W/W) PEG1000-17.0% (W/W) potassium phosphate ATPs at pH 9.2 with high-speed counter-current chromatograph at rotation speed of 850r/min and flow rate of 1.8mL/min, using upper phase as stationary phase. The purification recovery of ovalbumin was around 95%.

Original languageEnglish (US)
Pages (from-to)129-134
Number of pages6
JournalSheng wu gong cheng xue bao = Chinese journal of biotechnology
Volume21
Issue number1
StatePublished - Jan 1 2005
Externally publishedYes

Fingerprint

Egg White
Ovalbumin
Chromatography
Adenosine Triphosphate
Muramidase
Proteins
Myoglobin
Cytochromes
Liquid Chromatography
Phosphates
Equipment and Supplies
Research

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Purification of ovalbumin from hen egg white by high-speed counter-current aqueous two-phase chromatography. / Zhi, Wenbo; Deng, Qiu Yun; Song, Jiang Nan; Ouyang, Fan.

In: Sheng wu gong cheng xue bao = Chinese journal of biotechnology, Vol. 21, No. 1, 01.01.2005, p. 129-134.

Research output: Contribution to journalArticle

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