Studies on the Aggregate During the Refolding Procedure of Reduced-denatured Egg White Lysozymes by Protein Electrophoreses and Size-exclusion Chromatography

The aggregate of egg white lysozyme molecules formed during the refolding procedure of denatured-reduced egg white lysozymes was analyzed by protein electrophoreses and high-performance size-exclusion chromatography.When the denatured-reduced egg white lysozymes were renatured by dilution method,an observable amount of aggregate precipitate could be immediately formed.The results of lauryl sodium sulfate-polyacrylamide gel electrophoreses(SDS-PAGE) of the aggregate precipitate and supernatant and the result of high-performance size-exclusion chromatography of the supernatant indicated that,by wrongly linked intermolecular disulfide bonds soluble bi-molecular and tri-molecular egg white lysozyme aggregate could be simultaneously formed except being renatured to native and active egg white lysozymes during the refolding procedure of denatured-reduced egg white lysozyme;the aggregate precipitate could be further formed by the non-covalent bonds interaction between the soluble bi-molecular egg white lysozyme aggregates,and the soluble tri-molecular egg white lysozyme aggregate could still stay at the supernatant.