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

Yasunari Ono; Motohiro Kitajima; Shusaku Daikoku; Toshifumi Shiroya; Shoko Nishihara; Yoshimi Kanie; Katsuhiko Suzuki; Satoshi Goto; Osamu Kanie

doi:10.1039/B809316D

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Lab on a Chip

Issue 12, 2008

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

Yasunari Ono , Motohiro Kitajima , Shusaku Daikoku , Toshifumi Shiroya , Shoko Nishihara , Yoshimi Kanie , Katsuhiko Suzuki , Satoshi Goto and Osamu Kanie

Lab Chip, 2008, 8, 2168-2173

DOI: 10.1039/B809316D
Received 04 Jun 2008, Accepted 19 Aug 2008
First published on the web 16 Oct 2008

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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.

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Yasunari Ono
Motohiro Kitajima
Shusaku Daikoku
Toshifumi Shiroya
Shoko Nishihara
Yoshimi Kanie
Katsuhiko Suzuki
Satoshi Goto
Osamu Kanie

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

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