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Purification and molecular characterization of chitin deacetylase from Rhizopus nigricansJeraj, Nataša ...Chitin deacetylase is an enzyme, which catalyses the hydrolysis of N-acetamidobonds 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).Vir: Enzyme and microbial technology. - ISSN 0141-0229 (Letn. 39, 2006, str. 1294-1299)Vrsta gradiva - članek, sestavni delLeto - 2006Jezik - angleškiCOBISS.SI-ID - 21524441
Avtor
Jeraj, Nataša |
Kunič, Barbara |
Lenasi, Helena, 1945- |
Breskvar, Katja
Teme
Rhizopus |
Enzymology |
Cell Wall |
Chitin |
Metabolism |
Hydrolysis |
Chromatography, Deae-Cellulose |
Chromatography, Agarose |
Base Sequence |
Celična stena |
Hidroliza |
Hitin |
Kromatografija, DEAE-celuloza |
Rhizopus |
Baze, zaporedje |
Kromatografija, agaroza
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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Jeraj, Nataša | 21494 |
Kunič, Barbara | 15848 |
Lenasi, Helena, 1945- | 02934 |
Breskvar, Katja | 03079 |
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