fulltext.study @t Gmail

Galactosylation and sialylation of terminal glycan residues of human immunoglobulin G using bacterial glycosyltransferases with in situ regeneration of sugar-nucleotides

Paper ID Volume ID Publish Year Pages File Format Full-Text
18752 42741 2006 7 PDF Available
Title
Galactosylation and sialylation of terminal glycan residues of human immunoglobulin G using bacterial glycosyltransferases with in situ regeneration of sugar-nucleotides
Abstract

The N-glycans in human immunoglobulin G were engineered through the bacterial glycosyltransferase-catalyzed galactosylation and sialylation with in situ regeneration of sugar-nucleotides. For the galactosylation and sialylation, β1,4-galactosyltransferase from H. pylori and α2,3-sialyltransferase from N. gonorrhoeae were expressed in Escherichia coli. For the regeneration of UDP-galactose and CMP-NeuAc, the required enzymes were also expressed in E. coli and their extracts were employed as biocatalysts. Using the catalytic system, we carried out the galactosylation of terminal GlcNAc residues with UDP-Gal regeneration and sialylation of terminal Gal residues with CMP-NeuAc regeneration, respectively. When the oligosaccharides in human IgG were analyzed by HPLC (NP and WAX) and MALDI-TOF, the galactosylation reaction showed 86.1% increase in terminal galactosylation of N-linked oligosaccharide in IgG, and the sialylation reaction gave 69.6% increase in terminal sialylation. To achieve a maximum sialylation of IgG, simultaneous galactosylation and sialylation reaction was conducted by incubating all of the biocatalysts required for the galactosylation, sialylation and sugar-nucleotide regenerations in one reactor, resulting in 80.2% increase in terminal sialylation.

Keywords
N-linked oligosaccharide; Immunoglobulin G; β1,4-Galactosyltransferase; α2,3-Sialyltransferase; UDP-Gal; CMP-NeuAc
First Page Preview
Galactosylation and sialylation of terminal glycan residues of human immunoglobulin G using bacterial glycosyltransferases with in situ regeneration of sugar-nucleotides
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 39, Issue 1, 1 June 2006, Pages 60–66
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us