Specificity of chitosanase from Bacillus pumilus.
Partially (25-35%) N-acetylated chitosan was digested by chitosanase from Bacillus pumilus BN-262, and structures of the products, partially N-acetylated chitooligosaccharides, were analyzed in order to investigate the specificity of the chitosanase. The chitosanase produced glucosamine (GlcN) oligosaccharides abundantly, indicating that the chitosanase splits the beta-1,4-glycosidic linkage of GlcN-GlcN. The chitosanase also produced hetero-oligosaccharides consisting of glucosamine and N-acetyl-D-glucosamine (GlcNAc). Three types of the hetero-oligosaccharides purified by cation-exchange chromatography and HPLC were found to have GlcNAc residue at their reducing end and GlcN residue at their non-reducing end, indicating that the chitosanase can also split the linkage of GlcNAc-GlcN. The determination of the mode of action toward partially N-acetylated chitosan enables a classification of chitosanases according to their specificities and a more precise definition of chitosanases.
Fukamizo
T
T
Laboratory of Biophysical Chemistry, Faculty of Agriculture, Kinki University, Nara, Japan.
Ohkawa
T
T
Ikeda
Y
Y
Goto
S
S
D001407:Bacillus / Q000201:enzymology*; D002240:Carbohydrate Sequence; D002686:Chitin / Q000031:analogs & derivatives* / Q000378:metabolism; D048271:Chitosan; D005944:Glucosamine / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D017421:Sequence Analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013379:Substrate Specificity
D009844:Oligosaccharides; D002686:Chitin; D048271:Chitosan; D006026:Glycoside Hydrolases; C019868:chitosanase; D005944:Glucosamine