The intact cells of Rhizopus oligosporus NRRL2710, whose cell walls are abundant source of N-acetylglucosamine (GlcNAc) and glucosamine (GlcN), were digested with three chitinolytic enzymes, a GH-46 chitosanase from Streptomyces sp. N174 (CsnN174), a chitinase from Pyrococcus furiosus, and a chitinase from Trichoderma viride, respectively. Solubilization of the intact cells by CsnN174 was found to be the most efficient from solid state CP/MAS (13)C NMR spectroscopy. Chitosanase products from Rhizopus cells were purified by cation exchange chromatography on CM-Sephadex C-25 and gel-filtration on Cellulofine Gcl-25m. NMR and MALDI-TOF-MS analyses of the purified products revealed that GlcN-GlcNAc, (GlcN)2-GlcNAc, and (GlcN)2 were produced by the enzymatic digestion of the intact cells. The chitosanase digestion of Rhizopus cells was found to be an excellent system for the conversion of fungal biomass without any environmental impact.
Keywords: (GlcN)(n); 2-acetamido-2-deoxy-β-d-glucopyranose; 2-amino-2-deoxy-β-d-glucopyranose; CP/MAS (13)C NMR; Chitosanase; Fungal cell wall; GlcN; GlcNAc; MALDI-TOF-MS; NMR; Partially N-acetylated chitooligosaccharides; Rhizopus oligosporus; matrix assisted laser desorption/ionization time-of-flight mass spectrometry; nuclear magnetic resonance; β-1,4-linked oligosaccharide of GlcN with a polymerization degree of n.
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