Cloning and expression of a chitin deacetylase gene ( CDA2) from Saccharomyces cerevisiae in Escherichia coli: Purification and characterization of the cobalt-dependent recombinant enzyme

The chitin deacetylase gene CDA2 from Saccharomyces cerevisiae has been cloned and expressed in Escherichia coli and the recombinant enzyme purified to homogeneity and further characterized. The enzyme exhibits an apparent molecular mass of 35 kDa. When glycol chitin is used as substrate the optimum...

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Bibliographic Details
Published in:Enzyme and microbial technology 2003-05, Vol.32 (6), p.757-763
Main Authors: Martinou, Aggeliki, Koutsioulis, Dimitris, Bouriotis, Vassilis
Format: Article
Language:English
Subjects:
Biological and medical sciences
Chitin deacetylase
Escherichia coli
Fundamental and applied biological sciences. Psychology
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Saccharomyces cerevisiae
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Summary:The chitin deacetylase gene CDA2 from Saccharomyces cerevisiae has been cloned and expressed in Escherichia coli and the recombinant enzyme purified to homogeneity and further characterized. The enzyme exhibits an apparent molecular mass of 35 kDa. When glycol chitin is used as substrate the optimum temperature for enzyme activity is 50 °C and the pH optimum is 8.0. The enzyme requires at least two N-acetyl- d-glucosamine residues (chitobiose) for catalysis and is inhibited by acetate. The presence of CoCl 2 proved to be essential for enzyme activity. Seven amino acids could be eliminated from the C-terminus without a significant loss of enzyme activity. However, truncation of 12 or more amino acids from the N-terminus or 27 amino acids from the C-terminus resulted in complete loss of enzyme activity.
ISSN:0141-0229
1879-0909
DOI:10.1016/S0141-0229(03)00048-6