Characterization of Endoglucanase (F-I-III) Purified from Trichoderma sp. C-4

새로운 섬유소분해 균주 Trichoderma sp. C-4에서 분리한 Endoglucanase (F-I-III)에 대한 연구

  • Sul Ok Ju (Department of Biological Science, University of Ulsan) ;
  • Chung Dae Kyun (Institute and Department of Genetic Engineering, College of Natural Sciences, Kyung-Hee University) ;
  • Han In Seob (Department of Biological Science, University of Ulsan) ;
  • Jeong Choon Soo (Department of Biological Science, University of Ulsan)
  • 설옥주 (울산대학교 자연대학 생명과학부) ;
  • 정대균 (경희대학교 자연대학 유전공학과) ;
  • 한인섭 (울산대학교 자연대학 생명과학부) ;
  • 정춘수 (울산대학교 자연대학 생명과학부)
  • Published : 2005.03.01

Abstract

One of the endoglucanases, F-I-III, was purified from the culture filtrate of T. sp. C-4 through procedures including chromatography on Sephacryl S-200, DEAE-Sepharose A-50, and Chromatofocusing on Mono-P (FPLC). The molecular weight of the enzyme was determined to be about 56,000 Da by SDS-PAGE, and pI of 4.9 by analytical isoelectric focusing. F-I-III showed the highest enzyme activity at $55^{\circ}C$, and the pH optimum of the enzyme was 5.0. There was no loss of activity when the enzyme was incubated at $50^{\circ}C$ for 24 hours. The specific activity of the enzyme F-I-III toward the CMC was 315.4 U/mg. The Km value for $PNPG_2$ of F-I-III was 2.69 mM. N-terminal sequence of F-I-III was analyzed to be QPGTSTPEVHPKKLTTYK. It showed $95\%$ of homology to that of EGI from T. reesei. The presence of some metal ions (1 mM) had only a little effect on CMCase activity. The treatment of the reducing agents resulted in the increase of endoglucanase activity.

Keywords

C-4;characterization;protein sequencing;Trichoderma sp.

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