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

Yoshitake Maeda, Hidenori Yamada, Tadashi Ueda, Taiji Imoto

Research output: Contribution to journalArticle

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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
Publication statusPublished - May 1 1996

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Muramidase
Urea
Enzymes
Freezing
Melting point
Sarcosine
Temperature
Glucosamine
Differential Scanning Calorimetry
Ammonium Sulfate
Interchanges
Glycerol
Citric Acid
Disulfides
Glucose
Differential scanning calorimetry
Buffers

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Effect of additives on the renaturation of reduced lysozyme in the presence of 4 M urea. / Maeda, Yoshitake; Yamada, Hidenori; Ueda, Tadashi; Imoto, Taiji.

In: Protein Engineering, Vol. 9, No. 5, 01.05.1996, p. 461-465.

Research output: Contribution to journalArticle

Maeda, Yoshitake ; Yamada, Hidenori ; Ueda, Tadashi ; Imoto, Taiji. / Effect of additives on the renaturation of reduced lysozyme in the presence of 4 M urea. In: Protein Engineering. 1996 ; Vol. 9, No. 5. pp. 461-465.
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