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Overexpression and characterization of a novel chitinase gene from a marine bacterium Pseudomonas sp. BK1

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Title Overexpression and characterization of a novel chitinase gene from a marine bacterium Pseudomonas sp. BK1
 
Creator Jang, Moon-Sun
Lee, Young-Mi
Cho, Young-Su
Choi, Yong-Lark
Kim, Cherol-Ho
Lee, Young-Choon
 
Subject Pseudomonas
Chitinase
Cloning
Overexpression
Refolding
 
Description 339-344
The chitinase A (ChiA)-coding gene of Pseudomonas sp. BK1, which was isolated from a marine red alga Porphyra dentata, was cloned and expressed in Escherichia coli. The structural gene consists of 1602 bp encoding a protein of 534 amino acids, with a predicted molecular weight of 55,370 Da. The deduced amino acid sequence of ChiA showed low identity (less than 32%) with other bacterial chitinases. The ChiA was composed of multiple domains, unlike the arrangement of domains in other bacterial chitinases. Recombinant ChiA overproduced as inclusion bodies was solubilized in the presence of 8 M urea, purified in a urea-denatured form and re-folded by removing urea. The purified enzyme showed maximum activity at pH 5.0 and 40ºC. It exhibited high activity towards glycol chitosan and glycol chitin, and lower activity towards colloidal chitin. The enzyme hydrolyzed the oligosaccharides from (GlcNAc)₄ to (GlcNAc)₆, but not GlcNAc to (GlcNAc)3. The results suggest that the ChiA is a novel enzyme, with different domain structure and action mode from bacterial family 18 chitinases.
 
Date 2009-03-30T09:06:46Z
2009-03-30T09:06:46Z
2005-12
 
Type Article
 
Identifier 0301-1208
http://hdl.handle.net/123456789/3543
 
Language en_US
 
Relation C12N15/52; C12N9/28
 
Publisher CSIR
 
Source IJBB Vol. 42(6) [December 2005]