KR900702022A - 스테로이드를 생화학적으로 산화시키는 방법 및 이것을 위해 사용되도록 유전적으로 처리된 세포 - Google Patents

스테로이드를 생화학적으로 산화시키는 방법 및 이것을 위해 사용되도록 유전적으로 처리된 세포

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KR900702022A
KR900702022A KR1019900700022A KR900700022A KR900702022A KR 900702022 A KR900702022 A KR 900702022A KR 1019900700022 A KR1019900700022 A KR 1019900700022A KR 900700022 A KR900700022 A KR 900700022A KR 900702022 A KR900702022 A KR 900702022A
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expression cassette
heterologous dna
converting
protein
progesterone
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슬리즈쿠이스 헤르만
코넬리스 마리아 셀텐 게라더스
바스티안 스말 에렉
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안스 윌터 라벤
기스트-브로카데스 나암로제 베노오트하프
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Abstract

내용 없음

Description

스테로이드를 생화학적으로 산화시키는 방법 및 이것을 위해 사용되도록 유전적으로 처리된 세포
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 포유동물의 부신피질에서 일어나는 것과 같이 히드로코르티손내의 콜레스테롤 변환에 있어서의 연속단계에 포함되는 단백질에 대한 개요를 도시하고, 제2도는 플라스미드 pGBSCC-1의 구성을 도시하고, P4 5 0SCC- 서열 박스로 표시하며, 제3도는 합성유도(synlead)된 플라스미드 pTZ를 얻기 위해 플라스미드 pTZ 18R안에 5` - P4 5 0SCC- 서열을 함유하는 합성 PstI/HindⅢ 분획의 삽입을 도시한다.

Claims (25)

  1. 콜레스테롤을 히드로코르티손으로 변환시키기 위한 생화학적 경로에 있어서, 콜레스테롤을 프레그넨올론으로 변환시키는 단계; 프레그넨올론을 프로게스테론으로 변화시키는 단계; 프로게스테론을 17α-히드록시프로게스테론으로 변환시키는 단계; 17α-히드록시 프로게스테론을 코르텍솔론으로 변환시키는 단계; 코르텍솔론을 히드로코르티손으로 변환시키는 단계; 로 이루어지는 군으로부터 선택된 산화단계를 촉매하는데에 단독으로 서나 한가지 또는 그 이상의 부가 단백질과 함께 작용하는 단백질을 코드화하는 이종구조 DNA 코드서열, 및 재조합 숙주내에서 효과적인 대응 대조서열로 이루어지는 것을 특징으로 하고 상기 숙주에서 실시가능한 발현카세트.
  2. 제1항에 있어서, 이종구조 DNA 코드 서열이 제1항의 적어도 2개의 산화단계에서 단독으로 또는 하나 또는 그 이상의 부가단백질과 함께 촉매 작용을 하는 최소한 2가지 단백질을 코드화하는 것을 특징으로 하는 발현카세트.
  3. 제1항에 있어서, 발현카세트는 제1항의 산화단계의 군을 단독으로 또는 하나 또는 그 이상의 부가단백질과 함께 작용하는 단백질을 코드화하는 자체의 효과적 대응서열이 있는 부가적 이종구조 DNA가 적어도 한개 함유되어 있다는 것을 특징으로 하는 발현 카세트.
  4. 제1항 내지 3항중의 어느 한 항에 있어서, 상기 단백질이 곁사슬분리효소(P4 5 0SCC) ; 아드레노독신(ADX);아드레노독신 환원효소(ADR); β-히드록시스테로이드 탈수소효소/이소메라제(3β-HSD); 스테로이드-17α-히드록실라아제(P4 5 017α); NADPH 시토크롬 P4 5 0환원효소(RED); 스테로이드-21-히드록실라아제(P4 5 0C21), 및 스테로이드-11β-히드록실라아제(P4 5 011β)로 이루어지는 군으로부터 선택되는 것을 특징으로 하는 발현카세트.
  5. 제4항에 있어서, 상기 이종구조 DNA 코드서열이 소에서 유래하는 것을 특징으로 하는 발현카세트.
  6. 제4항 또는 제5항에 있어서, 상기 이종구조 DNA가 제4항의 군으로 부터 적어도 하나의 부가 단백질을 코드화 하는 것을 특징으로 하는 발현카세트.
  7. 제4항 또는 제5항에 있어서, 제3항의 군으로부터의 단백질을 코드화하는 자체의 효과적인 대조서열이 있는 적어도 하나의 부가적 이종구조 DNA가 함유되어 있는 것을 특징으로 하는 발현카세트.
  8. 제6항 또는 제7항에 있어서, 이종구조 DNA가 소의 P4 5 0SCC와 소의 ADX를 코드화하는 것을 특징으로 하는 발현카세트.
  9. 제5항에 있어서, 이종구조 DNA가 P4 5 0SCC 효소를 코드화하고 발현카세트는 pGBSCC-n 으로 표시되는 군으로부터 취해지며, 이때 n이 1부터 17까지의 정수인 것을 특징으로 하는 발현카세트.
  10. 제5항에 있어서, 이종구조 DNA가 P4 5 017α효소를 코드화하고 발현카세트는 pGB17α-n 으로부터 취해지며, 이때 n이 1부터 5까지의 정수인 것을 특징으로 하는 발현카세트.
  11. 제5항에 있어서, 이종구조 DNA는 P4 5 021C 효소를 코드화하고 발현카세트는 pGBC21-n 으로 표시되는 군으로부터 취해지며, 이때 n은 1부터 19까지의 정수인 것을 특징으로 하는 발현카세트.
  12. 제5항에 있어서, 이종구조 DNA가 P4 5 011β효소를 코드화하고 발현카세트는 pGB11β-n 으로 표시되는 군으로부터 취해지며, 이때 n이 1부터 4까지의 정수인 것을 특징으로 하는 발현카세트.
  13. 미생물, 식물 또는 동물의 세포로 이루어지고, 이종구조 DNA를 가지는 1항 내지 12항중의 어느 한 항에서 정의된 발현카세트가 함유되어 있는 것을 특징으로 하는 재조합 숙주세포 및 이것의 자세포.
  14. 제13항에 있어서, 숙주가 미생물인 것을 특징으로 하는 재조합 숙주세포 및 이것의 자세포.
  15. 제14항에 있어서, 숙주가 삭카로마이세스, 클뤼베로마이세스 또는 바실루스 종이거나 대장균인 것을 특징으로 하는 재조합 숙주세포 및 이것의 자세포.
  16. 제13 내지 15항중의 어느 한 항에서, 제1 내지 12항중의 어느 한 항에 정의된 발현카세트를 적어도 2개 함유하는 것을 특징으로 하는 재조합 숙주세포 및 이것의 자세포.
  17. 단백질이 형성되어 배양세포내에 축적될 수 있도록 하는 조건하에 영양배지내에서 재조합 세포를 배양하는 것으로 이루어지고 제13 내지 15항중의 어느 한 항에 정의된 바와같은 재조합 숙주세포인 것을 특징으로 하는 재조합 세포에 의해 외인성 단백질을 제조하는 방법.
  18. 효소가 형성되어 배양세포내에 축적될 수 있도록 하는 조건하에 영양배지내에서, 제16항에 정의된 재조합 숙주 세포에 의해 외인성 단백질혼합물을 제조하는 것을 특징으로 하는 방법.
  19. 배양액내에서 선택적 생화학적 산화반응 및 산화합물의 축적을 허용하는 조건에서 한가지 또는 그이상의 단백질 존재하에 산화되도록 화합물을 인큐베이션하고나서 산화된 화합물을 회수하는 것으로 이루어지고, 이때 단백질 또는 단백질들은 제17항 또는 18항의 방법에 의해 제조된 것을 특징으로 하는 실험상의 선택적 생화학적 산화방법.
  20. 원하는 산화반응이 일어나고 산화된 화합물이 배양액에 축적되는 조건에서 선택화합물의 존재하에 재조합세포를 배양하고 나서 산화화합물을 회수하는 것으로 이루어지며, 이때 재조합세포는 제13항 내지 16항중의 어느 한 항의 재조합숙주세포인 것을 특징으로 하는 선택적 화합물을 산화시키는 방법.
  21. 제19항 또는 제20항에 있어서, 산화반응이 스테롤 화합물의 곁사슬을 분리하여 프레그넨올론으로 제조하는 단계; 프레그넨올론을 프로게스테론으로 변환하는 단계; 프로게스테론을 17α-히드록시게스테론으로 변환하는 단계;17α-히드록시프로게스테론을 코르텍솔론으로 변환하는 단계; 및 크로텍솔론을 히드록코르티손으로 변환하는 단계; 로 이루어지는 군으로부터 선택되는 것을 특징으로 하는 방법.
  22. 제21항에 있어서, 산화반응이 스테롤화합물의 곁사슬을 분리하여 프레그넨올론을 형성시키는 것을 특징으로 하는 방법.
  23. 제21항에 있어서, 산화반응이 프로게스테론의 17α-히드록실화 반응인 것을 특징으로 하는 방법.
  24. 제21항에 있어서, 한 단계에서 동일 기질에 대하여 상기 군의 적어도 2가지 산화반응이 수행되는 것을 특징으로 하는 방법.
  25. 제19항 내지 24항중의 어느 한 항에 따라 제조된 활성화합물이 함유되어 있는 것을 특징으로 하는 제약 제조물.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019900700022A 1988-05-06 1989-05-08 스테로이드를 변환시키기 위한 다중유전자 시스템의 제조에 유용한 발현 카세트 KR100256025B1 (ko)

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CN1042567A (zh) * 1988-09-23 1990-05-30 吉斯特-布罗卡迪斯公司 类固醇多步氧化方法和所用的遗传工程细胞
EP0477961B1 (en) * 1990-09-26 1996-09-11 Sumitomo Chemical Company, Limited Mitochondrial P450
US5420027A (en) * 1991-01-10 1995-05-30 Board Of Regents, The University Of Texas System Methods and compositions for the expression of biologically active fusion proteins comprising a eukaryotic cytochrome P450 fused to a reductase in bacteria
US5240831A (en) * 1991-01-10 1993-08-31 Board Of Regents, The University Of Texas Methods and compositions for the expression of biologically active eukaryotic cytochrome p45os in bacteria
GB9615032D0 (en) 1996-07-17 1996-09-04 Univ Dundee Enzyme system
US9255256B2 (en) 1996-07-17 2016-02-09 Btg International Limited Expression of functional cytochorome P450 monooxygenase system in enterobacteria
FR2820145B1 (fr) * 2001-01-31 2004-01-23 Aventis Pharma Sa Souche de levure produisant des steroides de facon autonome
CN102272304B (zh) * 2009-01-07 2013-10-23 三菱化学株式会社 切断固醇侧链的酶蛋白质及其应用
JP6635917B2 (ja) * 2013-06-17 2020-01-29 サノフイＳａｎｏｆｉ チトクロムｐ４５０モノオキシゲナーゼ生体触媒作用のための全細胞系
EP3097113B1 (en) * 2014-01-20 2019-01-02 Sanofi Novel cytochrome p450 polypeptide with increased enzymatic activity
JP5800040B2 (ja) * 2014-01-29 2015-10-28 三菱化学株式会社 ステロール側鎖切断酵素蛋白質およびその利用

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NO904791L (no) 1991-01-04
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KR100256025B1 (ko) 2000-05-01
DK264890D0 (da) 1990-11-05
PT90484A (pt) 1989-11-30
WO1989010963A1 (en) 1989-11-16
AU3575989A (en) 1989-11-29
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JPH04500303A (ja) 1992-01-23
HUT54413A (en) 1991-02-28
NO904791D0 (no) 1990-11-05
DK175573B1 (da) 2004-12-13
CA1340616C (en) 1999-06-29
DE68929296D1 (de) 2001-06-21
JPH11308991A (ja) 1999-11-09
DK264890A (da) 1990-11-05
JP2963711B2 (ja) 1999-10-18
AU635494B2 (en) 1993-03-25

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