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Issue 12, 2008
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Sequential enzymatic glycosyltransfer reactions on a microfluidic device: Synthesis of a glycosaminoglycan linkage region tetrasaccharide

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Abstract

A microfluidic chip carrying three reaction chambers was designed and constructed to examine sequential multiple enzymatic reactions. The synthesis of oligosaccharides in living cells is carried out in the Golgi apparatus where multiple enzymes such as glycosidase and glycosyltransferases act on a variety of substrates to generate glycoconjugates that include glycolipids and glycoproteins. The regulatory mechanism of the process however remains unknown. A microchip-based analysis platform may provide a valuable tool with which to address the issue by mimicking the Golgi function. We thus examined 3 sequential glycosyltransfer reactions on a chip, and succeeded in the synthesis of a tetrasaccharide using immobilized enzymes. Also, the kinetic parameters for a recently identified glycosyltransferase, proteoglycan GalT-I, were obtained for the first time.

Graphical abstract: Sequential enzymatic glycosyltransfer reactions on a microfluidic device: Synthesis of a glycosaminoglycan linkage region tetrasaccharide

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Article information


Submitted
04 Jun 2008
Accepted
19 Aug 2008
First published
16 Oct 2008

Lab Chip, 2008,8, 2168-2173
Article type
Paper

Sequential enzymatic glycosyltransfer reactions on a microfluidic device: Synthesis of a glycosaminoglycan linkage region tetrasaccharide

Y. Ono, M. Kitajima, S. Daikoku, T. Shiroya, S. Nishihara, Y. Kanie, K. Suzuki, S. Goto and O. Kanie, Lab Chip, 2008, 8, 2168
DOI: 10.1039/B809316D

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