Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Regular Articles
Thermostability of Doubly Glycosylated Recombinant Lysozyme
Yoshio HASHIMOTOOsamu MUNEMURAKiyonari MASUMOTOTadashi UEDATaiji IMOTO
Author information
JOURNAL FREE ACCESS

2001 Volume 24 Issue 10 Pages 1102-1107

Details
Abstract
We prepared a lysozyme mutant (Q41S/R61S) introducing Asn-type glycosylation signal sites by yeast expression system. On purification by cation exchange column at pH 7, three fractions were obtained. Peptide mapping and mass-spectrometry showed the fractions were the derivatives glycosylated at both Asn39 and Asn59, at only Asn39, and not glycosylated. It was revealed that the processing of Asn-linked oligosaccharide at Asn39 and Asn59 occurred independently in yeast cells. The denaturation temperatures of these derivatives by differential scanning calorimetry were 76.0, 68.8, and 67.5°C at pH 3, respectively. The stabilization of glycosylated lysozyme depends on the degree of glycosylation. We concluded that stabilized proteins can be constructed by glycosylation at proper sites. Thermodynamic stabilization by the artificial double glycosylations on a protein has not yet been reported.
Content from these authors
© 2001 The Pharmaceutical Society of Japan
Previous article Next article
feedback
Top