A novel analytical method for D-glucosamine quantification and its application in the analysis of chitosan degradation by a minimal enzyme cocktail

Research output: Contribution to journalJournal article – Annual report year: 2016Researchpeer-review

  • Author: Mekasha, Sophanit

    Norwegian University of Life Sciences, Norway

  • Author: Toupalová, Hana

    Apronex s.r.o., Czech Republic

  • Author: Linggadjaja, Eka

    PT Biotech Surindo, Indonesia

  • Author: Tolani, Harish A.

    PT Biotech Surindo, Indonesia

  • Author: Anděra, Ladislav

    Apronex s.r.o., Czech Republic

  • Author: Arntzen, Magnus Ø.

    Norwegian University of Life Sciences, Norway

  • Author: Vaaje-Kolstad, Gustav

    Norwegian University of Life Sciences, Norway

  • Author: Eijsink, Vincent G. H.

    Norwegian University of Life Sciences, Norway

  • Author: Wittrup Agger, Jane

    Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Søltofts Plads, 2800, Kgs. Lyngby, Denmark

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Enzymatic depolymerization of chitosan, a β-(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 β-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).
Original languageEnglish
JournalCARBOHYDRATE RESEARCH
Volume433
Pages (from-to)18-24
ISSN0008-6215
DOIs
Publication statusPublished - 2016
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • Chitosan, Chitosanase, exo-β-glucosaminidase, Glucosamine, HPAEC-PAD

ID: 125637747