Journal of Korea Technical Association of The Pulp and Paper Industry (펄프종이기술)

Korea Technical Association of the Pulp and Paper Industry (한국펄프종이공학회)
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Purification of extracellular Cerrena unicolor laccase by means of affinity chromatography

친화크로마토그래프법을 이용한 Cerrena unicolor laccase의 정제특성

  • A. Leonowicz (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • L. Gianfreda (Department of chemical-Agricultural Sciences. University of Federico II, Via University) ;
  • J. Rogalski (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • M. Jaszek (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • M.W. Wasilewska (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • E. Malarczyk (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • A. Dawidowicz (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • M. Fink-Boots (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • G. Ginalska (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • Cho, Nam-Seok (Chungbuk National University)
  • Published : 1997.12.01
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Abstract

균체가 생산하는 laccase 효소가 다방면으로 이용되면서, 이 효소를 균체로부터 대량으로 생산하고 분리.정제하여야 할 필요성이 대두되고 있다. 아울러 이 효소의 활성을 유도하기 위한 inducer 로서 2,5-xylidine 이 주로 사용되고 있는바, 이 xylidine 의 유독성이 인정되면서 사람에게 독성을 주지 않는 환경친화적 inducer의 검색이 필요하게 되었다. 본 연구에서는 백색부후균 Cerrena unicolor가 분비하는 laccase 효소의 유도를 위한 inducer로서 ferulic acid를 사용하였으며, 균체로부터 생산 및 분리된 laccase 효소의 정제특성을 구명코자 하였다. 본효소(constitutive enzyme)로서 I 및 II를, ferulic acid를 inducer로 사용한 경우 inducing 효소 III을 분리.정제하였다. 본 효소 I 및 II의 Michaelis 정수는 각각 737 M, 716 M 이었고, inducing효소 III은 167 M 로서, 기질에 대한 높은 친화성을 보여주고 있다. 분자량도 각각 65 kD, 63 kD 였으며, inducing효소 III은 59 kD 였다. 두효소 모두 15-19%의 당 및 단백질분자당 4M의 동(Cu)을 함유하고 있었다. 정제효율은 효소 I 및 II가 10.1%, 9.4% 였으며, III은 3.2% 였다. 모든 효소의 최적 pH 는 5.5였으며, 최적온도는 비교적 높은 $40^{\circ}C$였다.

Keywords

References

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