한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제24권5호
  • /
  • Pages.567-573
  • /
  • 1996
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  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Bacillus licheniformis KFB-C14가 생산하는 내열성 Chitinase의 정제 및 특성

  • 홍범식 (고려대학교 생명공학원) ;
  • 윤호근 (고려대학교 생명공학원) ;
  • 신동훈 (고려대학교 생명공학원) ;
  • 조홍연 (고려대학교 생명공학원 및 연세대학교 생물산업소재연구센터)
  • 발행 : 1996.10.01
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초록

Chitinase (EC 3.2.1.14) from culture fluid of Bacillus licheniformis KFB-C14 was purified 66-folds to homogenity in overall yield of 21% by ammonium sulfate fractionation, DEAE-Toyopearl, Butyl-Toyopearl and TSK-Gel HW-55F column chromatography. The enzyme protein had a molecular weight of about 86,000 and was composed of one subunit. The enzyme was significantly stable not only at high temperature but also on treatment with organic solvents and protein denaturants such as SDS, urea and guanidine-HC1. The optimum temperature and pH for reaction was 60$\circ $C and 6.0, respectively. The enzyme activity was inhibited by only Mn$^{2+}$ ion, but not inhibited by EDTA, N- ethylmaleimide and pCMB. The enzyme had high activity with colloidal chitin (V$_{max}$: 421) and commercial chitin (V$_{max}$: 480), but not with typical substrates of exo type chitinase. The thermostable chitinase had an useful reactivity for producing functional chitooligosaccharide, showing the production of (GlcNAc)$_{1}, (GlcNAc)$_{3}$, and (GlcNAc)$_{2}$ as major product.

키워드

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