Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Characterization of Endoglucanase (F-II-II) Purified from Trichoderma sp. C-4

Trichoderma sp. C-4에서 분리한 endoglucanase(F-II-II)의 특성에 대한 연구

  • 설옥주 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 최지영 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 손영준 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 신지원 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 한인섭 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 정대균 (경희대학교 유전공학과 및 유전공학연구소) ;
  • 정춘수 (울산대학교 자연대학 생명과학부)
  • Published : 2000.03.01
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Abstract

One of endoglucanases(F-II-II) was purified from the culture filtrate of Trichoderma sp. C-4 through two step procedures including chromatography on Sephacryl S-200 and Sephacryl S-100. The molecular weight of the enzyme was determined to be about 26,000 by SDS-PAGE and the isoelectric point as 8.0 by analytical isoelectric focusing. The optimum temperature of the enzyme was $50^{\circ}C$ and the optimum pH was 5.0. No loss of activity was observed when the enzyme was preincubated at $50^{\circ}C$ for 24 hours. The specific activity of the enzyme toward carboxymethylcellulose (CMC) was estimated to be 776.2 U/mg. The internal amino acid sequence was analysed.

Trichoderma sp. C-4의 배양액으로부터 한 종류의 endoglucanase(F-II-II)를 Sephacryl S-200 및 Sephacryl S-100 chromatography를 통하여 분리하였다. 분리된 효소는 SDS-PAGE및 isoelectric focusing을 통하여 단일 band로 나타났으며, 분자량이 26,000, 등전점이 8.0으로 나타났다. 이 효소의 반응 최적온도와 최적 pH는 각각 $50^{\circ}C$, 5.0 이었으며, $50^{\circ}C$에서 24시간 동안 안정하였다. 분리된 효소의 carboxymethylcellulose에 대한 specific activity는 776.2 U/mg protein으로 나타났다. 이 효소의 아미노산 조성을 조사하였다. 효소를 trypsin으로 가수분해한 후 분해산물에 대한 단백질서열 분석을 행하였다. 그 결과 이 효소의 서열은 현재까지 밝혀진 다른 단백질과 homology를 갖지 않음이 확인되었다. 이 효소는 그 specific activity가 대단히 높고 아직 보고되지 않은 novel protein일 가능성이 대단히 높기 때문에 좋은 유전자 자원이 될 것으로 사료되었다.

Keywords

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