Journal
CARBOHYDRATE RESEARCH
Volume 433, Issue -, Pages 18-24Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carres.2016.07.003
Keywords
Chitosan; Chitosanase; Glucosamine; exo-beta-glucosaminidase; HPAEC-PAD
Funding
- European Union, ChiBio (Development of an integrated biorefinery for processing chitin rich biowaste to specialty and fine chemicals) [289284]
- Research Council of Norway, through the Marpol project [221576]
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Enzymatic depolymerization of chitosan, a beta-(1,4)-linked polycationic polysaccharide composed of D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) provides a possible route to the exploitation of chitin-rich biomass. Complete conversion of chitosan to mono-sugars requires the synergistic action of endo- and exo- chitosanases. In the present study we have developed an efficient and cost-effective chitosan-degrading enzyme cocktail containing only two enzymes, an endo-attacking bacterial chitosanase, ScCsn46A, from Streptomyces coelicolor, and an exo-attacking glucosamine specific beta-glucosaminidase, Tk-Glm, from the archaeon Thermococcus kodakarensis KOD1. Moreover, we developed a fast, reliable quantitative method for analysis of GlcN using high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The sensitivity of this method is high and less than 50 pmol was easily detected, which is about 1000-fold better than the sensitivity of more commonly used detection methods based on refractive index. We also obtained qualitative insight into product development during the enzymatic degradation reaction by means of ElectroSpray Ionization-Mass Spectrometry (ESI-MS). (C) 2016 Elsevier Ltd. All rights reserved.
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