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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Overexpression and characterization of thermostable chitinase from Bacillus atrophaeus SC081 in Escherichia coli

  • Cho, Eun-Kyung (Department of Food and Nutrition, College of Medical and Life Science, Silla University) ;
  • Choi, In-Soon (Department of Biological Science, College of Medical and Life Science, Silla University) ;
  • Choi, Young-Ju (Department of Food and Nutrition, College of Medical and Life Science, Silla University)
  • Received : 2010.11.01
  • Accepted : 2010.12.18
  • Published : 2011.03.31
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Abstract

The chitinase-producing strain SC081 was isolated from Korean traditional soy sauce and identified as Bacillus atrophaeus based on a phylogenetic analysis of the 16S rDNA sequence and a phenotypic analysis. A gene encoding chitinase from B. atrophaeus SC081 was cloned in Escherichia coli and was named SCChi-1 (GQ360078). The SCChi-1 nucleotide sequences were composed of 1788 base pairs and 596 amino acids, which were 92.6, 89.6, 89.3, and 78.9% identical to those of Bacillus subtilis (ABG57262), Bacillus pumilus (ABI15082), Bacillus amyloliquefaciens (ABO15008), and Bacillus licheniformis (ACF40833), respectively. A recombinant SCChi-1 containing a hexahistidine tag at the amino-terminus was constructed, overexpressed, and purified in E. coli to characterize SCChi-1. $H_6SCChi$-1 revealed a hydrolytic band on zymograms containing 0.1% glycol chitin and showed the highest lytic activity on colloidal chitin and acidic chitosan. The optimal temperature and pH for chitinolytic activity were $50^{\circ}C$ and pH 8.0, respectively.

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References

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