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, Shin Ichiro Shoda

Research output: Contribution to specialist publicationArticle

13 Citations (Scopus)

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-Glc-NAc), 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.

Original languageEnglish
Pages485-491
Number of pages7
Volume60
No.5
Specialist publicationHolzforschung
DOIs
Publication statusPublished - 2006 Aug 1

Keywords

  • Bacillus circulans WL-12
  • Chitinase A1
  • One-pot synthesis
  • Site-directed mutagenesis
  • Sugar oxazoline
  • Transglycosylation
  • Transition-state analog substrate

ASJC Scopus subject areas

  • Biomaterials

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