Expression, purification, and characterization of a cobalt-activated chitin deacetylase (Cda2p) from Saccharomyces cerevisiae.
Chitin deacetylase (Cda2p) (EC 3.5.1.41) from Saccharomyces cerevisiae has been purified from vegetative cells grown in galactose and further characterized. The enzyme is a glycoprotein with an apparent molecular mass of approximately 43 kDa and a carbohydrate content of approximately 18% by weight. With glycol chitin as substrate, the optimum temperature for enzyme activity is 50 degrees C and the pH optimum is 8.0. The enzyme requires at least two N-acetyl-D-glucosamine residues (chitobiose) for catalysis and is partially inhibited by acetate. Deglycosylation of the enzyme causes total loss of enzyme activity, which can be restored by the addition of COCl(2).
Martinou
Aggeliki
A
Enzyme Biotechnology Division, Institute of Molecular Biology and Biotechnology, Vassilika Vouton 711 10, Heraklion, Crete, Greece.
Koutsioulis
Dimitris
D
Bouriotis
Vassilis
V
D000581:Amidohydrolases / Q000235:genetics / Q000302:isolation & purification; D002686:Chitin / Q000031:analogs & derivatives / Q000096:biosynthesis; D048271:Chitosan; D003001:Cloning, Molecular; D015870:Gene Expression; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
D002686:Chitin; D048271:Chitosan; D000581:Amidohydrolases; C019866:chitin deacetylase