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Purification and molecular characterization of chitin deacetylase from Rhizopus nigricans

Paper ID Volume ID Publish Year Pages File Format Full-Text
18351 42719 2006 6 PDF Available
Title
Purification and molecular characterization of chitin deacetylase from Rhizopus nigricans
Abstract

Chitin deacetylase is an enzyme, which catalyses the hydrolysis of N-acetamido bonds of chitin, converting it to chitosan. In the present report we purified the chitin deacetylase from mycelial extracts of a zygomycete Rhizopus nigricans by sequential ammonium sulfate precipitation, CM Sepharose chromatography and DEAE-cellulose chromatography. The progress of enzyme purification was followed by measurement of enzyme activity using partially O-hydroxyethylated chitin (glycol chitin) radiolabelled in N-acetyl groups as a substrate. The apparent molecular mass of chitin deacetylase obtained by SDS-PAGE was approximately 100 kDa. A cDNA library containing a chitin deacetylase gene from R. nigricans was constructed and the complete gene was sequenced. The complete gene contains an open reading frame of 1341 nucleotides, which encodes a sequence of 447 amino acid residues. The estimated molecular mass is 47 kDa, suggesting that carbohydrate content is 53% by weight of the protein. The gene sequence consists of nucleotides encoding a conserved polysaccharide deacetylase domain located in the middle, covering 34% of the entire sequence. Overall, there were eight possible N-linked glycosylation sites. The deduced amino acid sequence shows the highest identity with chitin deacetylases from other zygomycetes Phycomyces blakesleeanus, Gongronella butleri, Rhizopus oryzae and Mucor rouxii (55%, 48%, 43% and 41% identity, respectively).

Keywords
cDNA; Cell wall; Chitin deacetylase; Enzyme purification; Rhizopus nigricans
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Purification and molecular characterization of chitin deacetylase from Rhizopus nigricans
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 39, Issue 6, 3 October 2006, Pages 1294–1299
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us