Efficient in vitro folding of the three‐disulfide derivatives of hen lysozyme in the presence of glycerol

FEBS Letters

Ohta, Katsuyuki²  Segawa, Shin-ichi¹  Tachibana, Hideki³  Sawano, Hiroshi³  Koumoto, Yasuko²  Sasaki, Yukio³ 

[1] Department of Physics, School of Science, Kwansei Gakuin University, Nishinomiya 662, Japan;Department of Biology, Faculty of Science, Kobe University, Rokkodai, Nada-ku, Kobe 657, Japan;Graduate School of Science and Technology Faculty of Science, Kobe University, Rokkodai, Nada-ku, Kobe 657, Japan

关键词: Protein folding;    Disulfide bond;    Lysozyme;    Renaturation;    Glycerol;    3SS-;    three disulfide bond-;   

DOI  :  10.1016/0014-5793(92)80466-T

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Four derivatives of hen lysozyme, each lacking one native disulfide bond of the four in authentic lysozyme, were produced in Escherichia coli by expressing synthetic mutant genes. In the reoxidation reaction of the reduced derivatives purified from inclusion bodies, the addition of glycerol significantly enhanced the efficiency of folding and ‘correct’ disulfide bond formation. This enabled simple chromatographical purification of refolded materials. Purified 3SS-derivatives all showed lytic activities and secondary structures comparable to authentic lysozyme, which directly showed that none of the four native disulfide bonds is a prerequisite for ‘correct’ in vitro folding.

【 授权许可】

Unknown