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).

Apronex s r o Nad Safinou 2 365 Vestec 252 42 Jesenice u Prahy Czech Republic

Department of Chemistry Biotechnology and Food Science Norwegian University of Life Sciences Ås Norway

Department of Chemistry Biotechnology and Food Science Norwegian University of Life Sciences Ås Norway; Center for Bioprocess Engineering Department of Chemical and Biochemical Engineering Technical University of Denmark Building 229 DK 2800 Kgs Lyngby Denmark

PT Biotech Surindo Komp Pelabuhan Perikanan Kejawanan Jl Pelabuhan Perikanan No 1 Cirebon West Java 45113 Indonesia

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