Characterization of the oligosaccharides assembled on the Pichia pastoris-expressed recombinant aspartic protease.

Abstract:

:Aspartic protease, widely used as a milk-coagulating agent in industrial cheese production, contains three potential N-glycosylation sites. In this study, we report the characterization of N-linked oligosaccharides on recombinant aspartic protease secreted from the methylotrophic yeast Pichia pastoris using a combination of mass spectrometric, 2D chromatographic, chemical and enzymatic methods. The carbohydrates from site I (Asn79) were found to range from Man6-17GlcNAc2 with 50% bearing a phospho-diester-motif, site II (Asn113) was not occupied and site III (Asn188) contained mostly uncharged species ranging from Man-13GlcNAc2. These charged groups are not affecting the transport through the secretion pathway of the recombinant glycoprotein. Changes from a molasses-based medium to a minimal salts-based medium led to a clear reduction of the degree of phosphorylation of the N-glycan population.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Montesino R,Nimtz M,Quintero O,García R,Falcón V,Cremata JA

doi

10.1093/glycob/9.10.1037

subject

Has Abstract

pub_date

1999-10-01 00:00:00

pages

1037-43

issue

10

eissn

0959-6658

issn

1460-2423

pii

cwc099

journal_volume

9

pub_type

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