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
Glycobiologyjournal_title
Glycobiologyauthors
Montesino R,Nimtz M,Quintero O,García R,Falcón V,Cremata JAdoi
10.1093/glycob/9.10.1037subject
Has Abstractpub_date
1999-10-01 00:00:00pages
1037-43issue
10eissn
0959-6658issn
1460-2423pii
cwc099journal_volume
9pub_type
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