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Glycosylation of human α 1 ‐antitrypsin in Saccharomyces cerevisiae and methylotrophic yeasts
Author(s) -
Kang Hyun Ah,
Sohn JungHoon,
Choi EuiSung,
Chung Bong Hyun,
Yu MeyongHee,
Rhee SangKi
Publication year - 1998
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19980315)14:4<371::aid-yea231>3.0.co;2-1
Subject(s) - saccharomyces cerevisiae , glycosylation , biochemistry , biology , pichia pastoris , asparagine , mannose , yeast , endoglycosidase h , glycoprotein , recombinant dna , heterologous expression , pichia , enzyme , gene , endoplasmic reticulum , golgi apparatus
Human α 1 ‐antitrypsin (α 1 ‐AT) is a major serine protease inhibitor in plasma, secreted as a glycoprotein with a complex type of carbohydrate at three asparagine residues. To study glycosylation of heterologous proteins in yeast, we investigated the glycosylation pattern of the human α 1 ‐AT secreted in the baker's yeast Saccharomyces cerevisiae and in the methylotrophic yeasts, Hansenula polymorpha and Pichia pastoris . The partial digestion of the recombinant α 1 ‐AT with endoglycosidase H and the expression in the mnn9 deletion mutant of S. cerevisiae showed that the recombinant α 1 ‐AT secreted in S. cerevisiae was heterogeneous, consisting of molecules containing core carbohydrates on either two or all three asparagine residues. Besides the core carbohydrates, variable numbers of mannose outer chains were also added to some of the secreted α 1 ‐AT. The human α 1 ‐AT secreted in both methylotrophic yeasts was also heterogeneous and hypermannosylated as observed in S. cerevisiae , although the overall length of mannose outer chains of α 1 ‐AT in the methylotrophic yeasts appeared to be relatively shorter than those of α 1 ‐AT in S. cerevisiae . The α 1 ‐AT secreted from both methylotrophic yeasts retained its biological activity as an elastase inhibitor comparable to that of α 1 ‐AT from S. cerevisiae , suggesting that the different glycosylation profile does not affect the in vitro activity of the protein. © 1998 John Wiley & Sons, Ltd.