Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Analysis of the Involvement of Chitin-Binding Domain of ChiCW in Antifungal Activity, and Engineering a Novel Chimeric Chitinase with High Enzyme and Antifungal Activities

  • Huang, Chien-Jui (Department of Plant Pathology and Microbiology, National Taiwan University) ;
  • Guo, Shu-Huei (Department of Plant Pathology and Microbiology, National Taiwan University) ;
  • Chung, Shu-Chun (Department of Plant Pathology and Microbiology, National Taiwan University) ;
  • Lin, Yu-Ju (Department of Plant Pathology and Microbiology, National Taiwan University) ;
  • Chen, Chao-Ying (Department of Plant Pathology and Microbiology, National Taiwan University)
  • Published : 2009.10.31
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Abstract

An antifungal chitinase, ChiCW, produced by Bacillus cereus 28-9 is effective against conidial germination of Botrytis elliptica, the causal agent of lily leaf blight. ChiCW as a modular enzyme consists of a signal peptide, a catalytic domain, a fibronectin type-III-like domain, and a chitin-binding domain. When two C-terminal domains of ChiCW were truncated, $ChiCW{\Delta}FC$ (lacking the chitin-binding domain and fibronectin type III-like domain) lost its antifungal activity. Since $ChiCW{\Delta}C$ (lacking the chitin-binding domain) could not be expressed in Escherichia coli as $ChiCW{\Delta}FC$ did, a different strategy based on protein engineering technology was designed to investigate the involvement of the chitin-binding domain of ChiCW ($ChBD_{ChiCW}$) in antifungal activity in this study. Because ChiA1 of Bacillus circulans WL-12 is a modular enzyme with a higher hydrolytic activity than ChiCW but not inhibitory to conidial germination of Bo. elliptica and the similar domain composition of ChiA1 and ChiCW, the C-terminal truncated derivatives of ChiA1 were generated and used to construct chimeric chitinases with $ChBD_{ChiCW}$. When the chitin-binding domain of ChiA1 was replaced with $ChBD_{ChiCW}$, the chimeric chitinase named ChiAAAW exhibited both high enzyme activity and antifungal activity. The results indicate that $ChBD_{ChiCW}$ may play an important role in the antifungal activity of ChiCW.

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