KR970703426A - 정제된 Myceliophthora 락카제 및 그것을 암호화 하는 핵산(PURIFIED MYCELIOPHTHORA LACCASES AND NUCLEIC ACIDS ENCODING SAME) - Google Patents

정제된 Myceliophthora 락카제 및 그것을 암호화 하는 핵산(PURIFIED MYCELIOPHTHORA LACCASES AND NUCLEIC ACIDS ENCODING SAME) Download PDF

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KR970703426A
KR970703426A KR1019960706887A KR19960706887A KR970703426A KR 970703426 A KR970703426 A KR 970703426A KR 1019960706887 A KR1019960706887 A KR 1019960706887A KR 19960706887 A KR19960706887 A KR 19960706887A KR 970703426 A KR970703426 A KR 970703426A
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laccase
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엠.베르카 란디
에이치. 브라운 스티븐
엑스유 펭
스크나이더 팔레
아니타 아아슬링 도리트
엠. 옥스엔뵐 카렌
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제임스 쉐한
노보 노르디스크 바이오테크 인코퍼레이티드
안네 제케르
노보 노르디스크 아크티에 셀스카브
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    • C12N9/0057—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10) with oxygen as acceptor (1.10.3)
    • C12N9/0061—Laccase (1.10.3.2)

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Abstract

본 발명은 Myceliophthora 락카제 암호화 서열을 함유하는 분리된 핵산 구조물과 그것에 의해 암호화된 락카제 단백질에 관한 것이다.

Description

정제된 Myceliophthora 락카제 및 그것을 암호화 하는 핵산(PURIFIED MYCELIOPHTHORA LACCASES AND NUCLEIC ACIDS ENCODING SAME)
[도면의 간단한 설명]
제1도는 pRaMB1에 있는 7.5EcoRI 단편의 제한지도를 도시한다. N.crassa 락카제 유전자 프로브에 혼성화 하는 영역은 어둡게 표시되어 있다.
제2도는 Myceliophthora thermophila 락카제의 뉴클레오티드(SEQ ID NO : 1) 및 아미노산(SEQ ID NO : 2)서열을 도시한다. 뉴클레오티드 서열중 소문자는 인트론의 위치를 가리킨다. 프로모터 영역중 추정상 TATA 및 CAAT 서열은 굵은 활자체로 하고 밑줄을 그었다. 공통 올가미 구조(PuCTPuAC)는 밑줄그어졌다.
제3도는 플라스미드 pRaMB5의 구조를 도시한다.
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (42)

  1. Myceliophthora 락카제를 암호화하는 서열을 함유하는 DNA 구조물.
  2. 제1항에 있어서, Myceliophthora thermophila 락카제를 암호화하는 서열로 이루어지는 것을 특징으로 하는 구조물.
  3. 제1항에 있어서, SEQ ID NO : 2에 도시된 아미노산 서열을 암호화하는 핵산서열로 이루어지는 것을 특징으로 하는 구조물.
  4. 제1항에 있어서, SEQ ID NO : 1에 도시된 핵산서열로 이루어지는 것을 특징으로 하는 구조물.
  5. 제1항에 있어서, NRRL B-21261에 함유된 핵산서열로 이루어지는 것을 특징으로 하는 구조물.
  6. 실질적으로 순수한 Myceliophthora 락카제 효소.
  7. 제6항에 있어서, Myceliophthora thermophila 락카제인 것을 특징으로 하는 효소.
  8. 제6항에 있어서, SEQ ID NO : 2에 도시된 아미노산 서열 또는 그것에 적어도 약 80% 상동성을 갖는 서열로 이루어지는 것을 특징으로 하는 효소.
  9. Myceliophthora 락카제를 암호화하는 서열을 함유하는 DNA구조물로 이루어지는 재조합 벡터.
  10. 제9항에 있어서, 구조물이 프로모터 서열에 작동가능하게 연결되는 것을 특징으로 하는 벡터.
  11. 제10항에 있어서, 프로모터는 균류 또는 효모 프로모터인 것을 특징으로 하는 벡터.
  12. 제11항에 있어서, 프로모터는 Aspergillus oryzae의 TAKA 아밀라제 프로모터인 것을 특징으로 하는 벡터.
  13. 제11항에 있어서, 프로모터는 Aspergillus niger 또는 Aspergillus awamori의 글루코 아밀라제(gluA) 프로모터인 것을 특징으로 하는 벡터.
  14. 제9항에 있어서, 또한 선택성 마커를 함유하는 것을 특징으로 하는 벡터.
  15. 제14항에 있어서, 선택성 마커는 amdS, pyrG, argB, niaD, sC 및 hygB로 구성된 군에서 선택되는 것을 특징으로 하는 벡터.
  16. 제14항에 있어서, 선택성 마커는 Aspergillus nidulans 또는 Aspergillus oryzae의 amdS마커, 또는 Aspergillus nidulans, Aspergillus niger, Aspergillus awamori 또는 Aspergillus oryzae의 pyrG마커인 것을 특징으로 하는 방법.
  17. 제14항에 있어서, Aspergillus oryzae의 TAKA 아밀라제 프로모터와 Aspergillus nidulans 또는 Aspergillus oryzae의 amdS 또는 pyrG마커 둘다로 이루어지는 것을 특징으로 하는 벡터.
  18. Myceliophthora 락카제를 암호화하는 핵산서열을 함유하는 비상동성 핵산 구조물로 이루어지는 재조합 숙주세포.
  19. 제18항에 있어서, 균류세포인 것을 특징으로 하는 세포.
  20. 제19항에 있어서, Aspergillus 세포인 것을 특징으로 하는 세포.
  21. 제18항에 있어서, 구조물이 숙주세포 게놈내로 통합되는 것을 특징으로 하는 세포.
  22. 제18항에 있어서, 구조물이 벡터상에 함유되는 것을 특징으로 하는 세포.
  23. 제18항에 있어서, SEQ ID NO : 2에 도시된 아미노산 서열을 암호화하는 서열을 함유하는 구조물로 이루어지는 것을 특징으로 하는 세포.
  24. 락카제 효소를 얻기 위한 방법에 있어서, 그 효소의 발현에 기여하는 조건하에 Myceliophthora 락카제 효소를 암호화하는 핵산서열을 함유하는 DNA 구조물로 이루어진 재조합 숙주세포를 배양하고 그 배양물로부터 효소를 회수하는 것으로 이루어지는 방법.
  25. 제24항의 방법에 의해 얻어진 Myceliophthora 효소.
  26. 최적 pH에서 시링알다진에 대해 적어도 30 SOU/㎎의 비활성을 가지는 자낭균류 또는 불완전균류 락카제.
  27. 활성 재조합 구리-함유 효소의 수율을 향상시키는 방법에 있어서, 적어도 약 0.02mM구리의 존재하에 그 효소의 발현에 기여하는 조건하에 구리함유 효소를 암호화하는 서열을 함유하는 DNA구조물로 이루어진 재조합 숙주세포를 배양하는 것으로 이루어지는 방법.
  28. 용액에서 리그닌 또는 리그노술페이트 기질을 중합하기 위한 방법에 있어서, 기질을 Myceliophthora 락카제와 접촉시키는 것으로 이루어지는 방법.
  29. Krafe펄프에서 원위치 해중합을 위한 방법에 있어서, 펄프를 Myceliophthora 락카제와 접촉시키는 것으로 이루어지는 방법.
  30. 염료 또는 염료 전구체를 산화시키기 위한 방법에 있어서, 염료를 Myceliophthora 락카제와 접촉시키는 것으로 이루어지는 방법.
  31. 머리를 염색하기 위한 방법에 있어서, 염료전구체에서 염료로의 산화를 허용하기에 충분한 조건하에 잠시 적어도 한가지 변형제의 존재 또는 부재하에 Myceliophthora 락카제를 적어도 한가지 염료전구체와 접촉시키는 것으로 이루어지는 방법.
  32. 제31항에 있어서, 염료전구체는 디아민, 아미노페놀 및 페놀로 구성된 군에서 선택되는 것을 특징으로 하는 방법.
  33. 제31항에 있어서, 변형제가 사용될때 변형제는 메타-디아민, 메타-아미노페놀 또는 폴리페놀인 것을 특징으로 하는 방법.
  34. 제32항에 있어서, 염료전구체는 오르토-또는 파라-디아민 또는 아미노페놀로 구성된 군에서 선택된 1차 중간체인 것을 특징으로 하는 방법.
  35. 제31항에 있어서, 한가지 보다 많은 염료가 사용되는 것을 특징으로 하는 방법.
  36. 제31항에 있어서, 한가지 보다 많은 염료가 사용되는 것을 특징으로 하는 방법.
  37. 제31항에 있어서, 1차중간체 및 변형제 둘다가 사용되는 것을 특징으로 하는 방법.
  38. 적어도 하나의 염료전구체와 조합된 Myceliophthora 락카제로 이루어지는 염료 조성물.
  39. 적어도 하나의 1차 중간체 및 적어도 하나의 변형체와 조합된 Myceliophthora 락카제로 이루어지는 염료 조성물.
  40. 무산소 분위기에서 Myceliophthora 락카제 및 적어도 하나의 염료전구체로 이루어진 염료 조성물을 함유하는 용기.
  41. 제40항에 있어서, 적어도 하나의 변형제와 조합된 적어도 하나의 1차중간체 염료전구체를 함유하는 것을 특징으로 하는 용기.
  42. 페놀계 또는 아닐린 화합물을 중합 또는 산화시키는 방법에 있어서, 페놀계 또는 아닐린 화합물을 Myceliophthora 락카제와 접촉시키는 것으로 이루어지는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019960706887A 1994-06-03 1995-05-31 정제된 Myceliophthora 락카제 및 그것을 암호화 하는 핵산(PURIFIED MYCELIOPHTHORA LACCASES AND NUCLEIC ACIDS ENCODING SAME) KR970703426A (ko)

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US25378194A 1994-06-03 1994-06-03
US44114695A 1995-05-15 1995-05-15
US08/253,781 1995-05-15
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PCT/US1995/006815 WO1995033836A1 (en) 1994-06-03 1995-05-31 Phosphonyldipeptides useful in the treatment of cardiovascular diseases

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