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Efficient in vitro folding of the three‐disulfide derivatives of hen lysozyme in the presence of glycerol
Author(s) -
Sawano Hiroshi,
Koumoto Yasuko,
Ohta Katsuyuki,
Sasaki Yukio,
Segawa Shin-ichi,
Tachibana Hideki
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80466-t
Subject(s) - lysozyme , disulfide bond , folding (dsp implementation) , chemistry , glycerol , in vitro , muramidase , biochemistry , inclusion bodies , escherichia coli , protein folding , lytic cycle , combinatorial chemistry , biology , gene , virus , virology , electrical engineering , engineering
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.