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Enzyme-Catalyzed Synthesis of a Hybrid N-Linked Oligosaccharide using N-Acetylglucosaminyltransferase I

✍ Scribed by Rui Chen; Mark A. Pawlicki; Brian S. Hamilton; Thomas J. Tolbert

Book ID: 101421287
Publisher: John Wiley and Sons
Year: 2008
Tongue: English
Weight: 224 KB
Volume: 350
Category: Article
ISSN: 1615-4150
DOI: 10.1002/adsc.200800265

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✦ Synopsis

Abstract

The soluble catalytic domain of human N‐acetylglucosaminyltransferase I was purified from Escherichia coli and utilized in the enzyme‐catalyzed conversion of high mannose N‐linked oligosaccharide 1 into the rare hybrid oligosaccharide 2. Analysis of the reaction showed that the conversion of high mannose 1 into hybrid oligosaccharide 2 proceeded to 100% completion as assessed by MALDI‐TOF‐MS. Purification of the large polar oligosaccharide by gel filtration and silica gel chromatography afforded a 42% isolated yield of oligosaccharide 2. This enzyme‐catalyzed reaction can be utilized to produce rare hybrid oligosaccharides for biochemical and structural studies.

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