Purification and Characteristics of Chitosanase from Bacillus sp. HW-002

  • Lee , Hyean-Woo (Department of Biochmistry, Wonju College of Medicine, Yonsei University) ;
  • Choi, Jong-Whan (Department of Biochmistry, Wonju College of Medicine, Yonsei University) ;
  • Han, Dong-Pyou (Department of Biochmistry, Wonju College of Medicine, Yonsei University) ;
  • Park, Myoung-Jin (Department of Microbiological Engineering, College of Engineering, Konkuk University) ;
  • Lee, No-Woon (Department of Microbiological Engineering, College of Engineering, Konkuk University) ;
  • Yi, Dong-Heui (Department of Microbiological Engineering, College of Engineering, Konkuk University)
  • Published : 1996.02.01

Abstract

Chitosanase from Bacillus sp. HW-002 was purified with CM-cellulose column chromatography, and HPLC with DEAE- TSK gel and YMC-pack Diol 120. The purified enzyme appeared as a single band on SDS-polyacrylamide gel. The molecular weight of the enzyme was estimated to be about 46 kDa on SDS-polyacrylamide gel, and was estimated to be about 23 kDa by GFC. The optimal pH of chitosanolytic activity was about pH 5.5-6.0, and the purified enzyme was most stable at pH 5.0. The optimal temperature of chitosanolytic activity was $65^{\circ}C$ and the enzyme was stable at $45^{\circ}C$ for 1 h. Chitosan was the most favorable substrate among various $\beta$-glucan. UVmax of the purified enzyme was 195 nmand was not noted around 280 nm. The main product of enzyme reaction with chitosan was chitobiose.

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