title: Purification and characterization of chitin deacetylase from Colletotrichum lindemuthianum.
abstract: Chitin deacetylase (EC 3.5.1.41), the enzyme that catalyzes the hydrolysis of acetamido groups of N-acetyl-D-glucosamine in chitin, has been purified to homogeneity from the culture filtrate of the fungus Colletotrichum lindemuthianum and further characterized. The enzyme is a glycoprotein, and its apparent molecular mass was determined to be approximately 150 kDa. The glycosylation pattern of the enzyme is consistent with a mixture of N-linked glycans including oligomannosidic hybrid and/or complex type, and its carbohydrate content is approximately 67% by weight. Chitin deacetylase is active on several chitinous substrates and chitin derivatives, is not considerably inhibited by carboxylic acids, especially acetic acid, and exhibits a remarkable thermostability. The enzyme requires at least two N-acetyl-D-glucosamine residues (chitobiose) for catalysis. When glycol chitin (a water-soluble chitin derivative) was used as substrate, the optimum temperature for enzyme activity was determined to be 50 degrees C, and the optimum pH was approximately 8.5.
authors.affiliation: Enzyme Technology Division, University of Crete, Greece.
authors.forename: I
authors.initials: I
authors.lastname: Tsigos
authors.forename: V
authors.initials: V
authors.lastname: Bouriotis
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003386:Brachyura; D002241:Carbohydrates / Q000032:analysis; D002686:Chitin / Q000378:metabolism; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D033364:Decapoda; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005658:Fungi / Q000201:enzymology*; D007700:Kinetics; D009005:Monosaccharides / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D009005:Monosaccharides; D009844:Oligosaccharides; D002686:Chitin; D000581:Amidohydrolases; C019866:chitin deacetylase