Functional Properties of Glycosylated Lysozyme Secreted in Pichia pastoris(Food & Nutrition Sience)
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概要
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Various mutant lysozymes having the N-glycosylation signal sequence, R21T (Asn^<19>-Tyr^<20>-Thr^<21>), G49N (Asn^<49>-Ser^<50>-Thr^<51>), R21T/G49N (Asn^<19>-Tyr^<20>-Thr^<21>/Asn^<49>-Ser^<50>-Thr^<51>), were secreted in the Pichia pastoris expression system. The secreted amounts of these mutant glycosylated lysozymes were almost the same as those of wild-type lysozyme (about 30 mg/liter). Glycosylation of the mutant lysozymes was confirmed by SDS-PAGE patterns, Endo-H treatment, TOF-MS analysis and chemical analysis. The composition of the carbohydrate chain attached to the single glycosylated lysozymes, R21T and G49N, was GlcNAc_2Man_<9-11>, while that of the double glycosylated lysozyme, R21T/G49N, was GlcNAc_4Man_<27-32>. The results of a CD analysis and lytic activity suggested that the conformation of the single glycosylated lysozymes had been conserved, while that of the double glycosylated lysozyme was less stable. The emulsifying properties of the lysozyme when glycosylated were greatly improved, being especially noteworthy in the double glycosylated lysozyme.
- 2003-11-23
著者
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SAITO AKIRA
Department of Pediatrics, Hirosaki University School of Medicine
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USUI Masakatsu
Department of Food Science and Technology, National Fisheries University
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Azakami Hiroyuki
Department of Fermentation Technology, Faculty of Engineering, Hiroshima University
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KATO Akio
Department of Agricultural Chemistry, Yamaguchi University
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Kato A
The Institute For Bio-medical Research Teijin Limited
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Usui Masakatsu
Department Of Biological Chemistry Yamaguchi University
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SONG Youtao
Department of Biological Chemistry, Yamaguchi University
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SAKO Yukikazu
Department of Biological Chemistry, Yamaguchi University
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Song Youtao
Department Of Biological Chemistry Yamaguchi University
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Azakami H
Department Of Biotechnology Graduate School Of Engineering Osaka University:school Of Dentistry Toku
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Azakami Hiroyuki
Department Of Biological Chemistry Faculty Of Agriculture Yamaguchi University
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Sako Yukikazu
Department Of Biological Chemistry Yamaguchi University
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Kato A
Department Of Biological Chemistry Yamaguchi University
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Saito A
Department Of Biological Chemistry Yamaguchi University
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Kato Akio
Department Of Agricultural Chemistry Faculty Of Agriclture Yamaguchi University
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Saito Akira
Department Of Pathology Juntendo University Urayasu Hospital
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Saito Akira
Department of Biological Chemistry, Yamaguchi University
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Saito Akira
Department of Applied and Bioapplied Chemistry, Graduate School of Engineering, Osaka City University
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