Thermostability of Doubly Glycosylated Recombinant Lysozyme.
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- HASHIMOTO Yoshio
- Graduate School of Pharmaceutical Sciences, Kyushu University
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- MUNEMURA Osamu
- Graduate School of Pharmaceutical Sciences, Kyushu University
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- MASUMOTO Kiyonari
- Graduate School of Pharmaceutical Sciences, Kyushu University
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- UEDA Tadashi
- Graduate School of Pharmaceutical Sciences, Kyushu University
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- IMOTO Taiji
- Graduate School of Pharmaceutical Sciences, Kyushu University
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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.
Journal
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- Biological and Pharmaceutical Bulletin
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Biological and Pharmaceutical Bulletin 24 (10), 1102-1107, 2001
The Pharmaceutical Society of Japan
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Details 詳細情報について
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- CRID
- 1390001204624726272
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- NII Article ID
- 110003638364
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- NII Book ID
- AA10885497
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- COI
- 1:CAS:528:DC%2BD3MXnt12jtLs%3D
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- ISSN
- 13475215
- 09186158
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- NDL BIB ID
- 5926460
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- PubMed
- 11642311
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed