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Stepwise synthesis of chitooligosaccharides through a transition-state analogue substrate catalyzed by mutants of chitinase A1 from Bacillus circulans WL-12

  • Michinari Kohri , Atsushi Kobayashi , Masato Noguchi , Shin-ichi Kawaida , Takeshi Watanabe and Shin-ichiro Shoda
Published/Copyright: August 16, 2006
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Holzforschung
From the journal Volume 60 Issue 5

Abstract

Several mutants of chitinase A1 from Bacillus circulans WL-12 have been prepared by site-directed mutagenesis of tryptophan 433. The hydrolytic activity of these mutants toward p-nitrophenyl chitobioside was lower than that of wild-type chitinase A1. The mutants were found to catalyze the transglycosylation reaction of the 1,2-oxazoline derivative of N-acetyllactosamine (Gal-GlcNAc-oxa) to N,N′′-diacetylchitobiose (GlcNAc-GlcNAc), giving rise to a transglycosylated tetrasaccharide product (Gal-GlcNAc-GlcNAc-GlcNAc) in high yield. Based on these results, a one-pot synthesis of chitooligosaccharides by combined use of mutant W433Y and β-galactosidase has been demonstrated. By controlling the reaction conditions, chitotriose and chitotetraose were obtained in 74% and 40% yield (determined by HPLC), respectively, without isolating any intermediate products.

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Corresponding author. Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aoba 6-6-11-514, Aoba-ku, Sendai 980-8579, Japan

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Published Online: 2006-08-16
Published in Print: 2006-08-01

©2006 by Walter de Gruyter Berlin New York

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