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. 1997 Dec 15;328(Pt 3):945–949. doi: 10.1042/bj3280945

Serratia marcescens chitobiase is a retaining glycosidase utilizing substrate acetamido group participation.

S Drouillard 1, S Armand 1, G J Davies 1, C E Vorgias 1, B Henrissat 1
PMCID: PMC1219008  PMID: 9396742

Abstract

The stereochemistry of the reaction catalysed by Serratia marcescens chitobiase was determined by HPLC separation of the anomers of N-acetylglucosamine produced during the hydrolysis of p-nitrophenyl N-acetyl-beta-d-glucosaminide (PNP-GlcNAc). In the early stages of the reaction, the beta-anomer was found to prevail, whereas the alpha-anomer dominated at mutarotation equilibrium. This established that chitobiase hydrolyses glycosidic bonds with overall retention of the anomeric configuration. Chitobiase-catalysed hydrolysis of PNP-GlcNAc was competitively inhibited by a series of chito-oligosaccharides (degree of polymerization 2-5) that were selectively de-N-acetylated at their non-reducing end. The results are in accord with the participation of the acetamido group at C-2 of the substrate in the catalytic mechanism of chitobiase and related enzymes.

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Selected References

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