Effect of additives on the renaturation of reduced lysozyme in the presence of 4 M urea

Yoshitake Maeda, Hidenori Yamada, Tadashi Ueda, Taiji Imoto

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Abstract

Reduced lysozyme was renatured by sulfhydryl-disulfide interchange reactions at pH 8.0 in the presence of 4 M urea, with or without additives at 40°C. In the absence of additives, the final folding yield of reduced lysozyme was ~40%. In the presence of sarcosine, glycerol, ammonium sulfate, N-acetyl glucosamine and glucose, its folding yields increased in all cases. In particular, yields increased up to 90% in the presence of 4 M sarcosine. On the other hand, the melting temperatures of lysozyme with or without additives in 0.02 M citrate buffer (pH 6.0) were evaluated using differential scanning calorimetry. In the absence of additive, the melting temperature of lysozyme was 73.8°C. In the presence of additives, all melting temperatures were higher than that of lysozyme in the absence of additives. Moreover, there was a good correlation on addition of additives between an increase in the folding yield of reduced lysozyme with 4 M urea and an increase in the melting temperature without 4 M urea. Therefore, we conclude that additives, which stabilize native lysozyme, are effective at increasing the folding yield of reduced lysozyme in 4 M urea.

Original languageEnglish
Pages (from-to)461-465
Number of pages5
JournalProtein Engineering
Volume9
Issue number5
DOIs
Publication statusPublished - May 1996

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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