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Cho, Moo Je
Ulsan National Institute of Science and Technology
Research Interests
  • Calcium Signaling

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Isolation and characterization of the 54-kDa and 22-kDa chitinase genes of Serratia marcescens KCTC2172

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Title
Isolation and characterization of the 54-kDa and 22-kDa chitinase genes of Serratia marcescens KCTC2172
Author
Gal, SWChoi, JYKim, CYCheong, YHChoi, YJBahk, JDLee, SYCho, Moo Je
Keywords
Chitinase; Cloning; Serratia marcescens
Issue Date
199706
Publisher
WILEY-BLACKWELL
Citation
FEMS MICROBIOLOGY LETTERS, v.151, no.2, pp.197 - 204
Abstract
A DNA fragment (pCHI54225 containing two genes encoding a 54-kDa and a 22-kDa chitinase was isolated from a cosmid DNA library of Serratia marcescens KCTC2172. The complete nucleotide sequence of pCHI5422 consisting of 4581 bp was determined. The nucleotide sequence of the 22-kDa chitinase consists of 681 bp of open reading frame encoding 227 amino acids and is located 1422 bp downstream of the translation termination codon of the 54- kDa chitinase sequence. The 54-kDa chitinase gene consisted of 1497 bp in a single open reading frame encoding 499 amino acids. The genes encoding the 54-kDa and 22-kDa chitinase were separately subcloned in Escherichia coli and the individual chitinases were expressed and purified from the culture broth using chitin affinity chromatography. When chitohexaose was used as substrate, the major product of the enzymatic reaction of both the 54-kDa and 22-kDa chitinases was a (GlcNAc)2 dimer with a minor amount of monomer. The specific activity of the 54-kDa and 22-kDa chitinases were 300 μM (min)-1 mg-1 and 17 μM (min)-1 mg-1 on the natural swollen chitin, respectively.
URI
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DOI
http://dx.doi.org/10.1016/S0378-1097(97)00159-6
ISSN
0378-1097
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