KR920701449A - 온도, 산 및/또는 알칼리에 대한 안정성이 증가된 변이체 미생물의 α-아밀라제 - Google Patents

온도, 산 및/또는 알칼리에 대한 안정성이 증가된 변이체 미생물의 α-아밀라제

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KR920701449A
KR920701449A KR1019910700227A KR910700227A KR920701449A KR 920701449 A KR920701449 A KR 920701449A KR 1019910700227 A KR1019910700227 A KR 1019910700227A KR 910700227 A KR910700227 A KR 910700227A KR 920701449 A KR920701449 A KR 920701449A
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amylase
variant
gene
starch
expression
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KR1019910700227A
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KR0165550B1 (ko
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요하네스 크박 빌헬무스
라로체 이페스
빌헬무스 헤르마너스 플레브레흐트 아드리아누스
스탄센스 파트리크
라우베리즈 마크
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기스트-브로카데스 나암로제 베노오트하프
플랜드 제네틱 시스템스 나암로제 베노오트하프
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
    • C12N15/75—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Bacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411—Amylases
    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411—Amylases
    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source

Abstract

내용 없음

Description

온도, 산 및/또는 알칼리에 대한 안정성이 증가된 변이체 미생물의 α-아밀라제
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 pMa 5-8의 뉴클레오티드 서열임.

Claims (21)

  1. α-아밀라제를 코드하는 변이된 DNA서열의 발현물인 변이체 α-아밀라제에 있어서, 야생형 효소와는 적어도 하나 이상의 아미노산이 다른 아미노산 서열을 가지고, 상기 변이체 α-아밀라제는 전분의 분해 및/또는 직물 풀빼기에 이용되는, 아미노산의 치환에 기인하는 개선된 특성을 나타내는 것을 특징으로 하는 변이체 α-아밀라제.
  2. 제1항에 있어서, 개선된 열안전성을 나타내는 것을 특징으로 하는 α-아밀라제.
  3. 제1항에 있어서, 6.5이하 및/또는 7.5이상의 pH에서 개선된 안정성을 나타내는 것을 특징으로 하는 α-아밀라제.
  4. 제1항에 있어서, 개선된 열안정성 및 산안정성을 나타내는 것을 특징으로 하는 α-아밀라제.
  5. 제1내지 4항중의 어느 한 항에 있어서, 변이체 효소가 유래한 원 유전자가 미생물 바람직하게는 바실루스(Bacillus)균주으로 부터 얻어지는 것을 특징으로 하는 α-아밀라제.
  6. 제5항에 있어서, 상기 유전자가 B. 스테아로더모필루스(B. stearothermophilus),B. 리케니포미스(B. licheniformis)및 B. 아밀로리퀴에파시엔스(B. amyloliquefaciins)로 구성되는 군으로 부터 선택된 균주의 야생형 유전자로 부터 유도된 것을 특징으로 하는 α-아밀라제.
  7. 제6항에 있어서, 상기 α-아밀라제가 111.133 및 149위치 또는 상동 a-아밀라제의 대응 위치에 하나 또는 그 이상의 아미노산이 치환됨으로써 바실루스 리케미포미스(Bacillus licheniformis)로 부터 얻어진 야생형 α-아밀라제와 다른 것을 특징으로 하는 α-아밀라제.
  8. 제7항에 있어서, 다음과 같은 아미노산 치환체 : Ala-111-Thr, His-133-Tyr, Yhr-149-ILe를 하나 또는 그 이상 포함하는 것을 특징으로 하는 α-아밀라제.
  9. 제1내지 8항중 어느 한 항에 있어서, 정의된 α-아밀라제를 코드하고 있는 변이 유전자.
  10. 제9항의 변이 유전자로 이루어진 것을 특징으로 하는 발현 벡터.
  11. 제10항의 발현 벡터를 수용하는 숙주세포.
  12. 형질전환전에 세포의 전분분해 효소를 실질적으로 생산할 수 없는 숙주세포로서, 제10항의 발현 벡터로써 형질전환되는 것을 특징으로 하는 숙주세포.
  13. 제12항에 있어서, B. 리케니포미스 T9(B. licheniformis T9)인 것을 특징으로 하는 숙주세포.
  14. E. coil에서 클론 유전자의 발현이 구조 유전자로부터 조절 서열의 물리적 분리에 의하여 불가능하고, 바실루스(Bacillus)에서 클로 유전자의 발현이 단일 제한 효소에 의한 절단 및 뒤이은 재환형화에 의해 회복되는 것을 특징으로 하는 바실루스/E. 콜리(Bacillus/E. Coli)셔틀벡터.
  15. 전분분해 효소 또는 그단편을 코드하는 클론 유전자를 변이시키고, 얻어진 변이 아밀라제 유전자 또는 유전자군을 단리하고, 상기 변이 아밀라제 유전자 또는 유전자군의 발현 및 생산을 위하여 적당한 숙주 균주에 도입하고, 생산된 변이 아밀라제를 회수하고 및 전분분해 또는 직물 풀빼기에 이용되는 개선된 특성을 가지는 이 변이 아밀라제를 동정하는 방법으로 이루어지는 것을 특징으로 하는 전분분해 또는 직물 풀빼기에 이용되는 개선된 특성을 가진 전분분해 효소의 제조방법.
  16. 제11내지 13항중 어느 한 항에 따른 숙주세포를 적당한 배지에서 배양하고, 생산된 α-아밀라제를 회수하는 것으로, 이루어지는 것을 특징으로 하는 변이 α-아밀라제의 제조방법.
  17. 전분분해 및 직물 풀빼기에서 제1내지 8항중 어느 한 항에 따른 α-아밀라제의 사용.
  18. 제1내지 8항중 어느 한 항에 따른 변이 α-아밀라제의 사용으로 이루어진 것을 특징으로 하는 전분 분해 방법.
  19. 제1내지 8항중 어느 한 항에 따른 변이 α-아밀라제의 사용으로 이루어지는 것을 특징으로 하는 직물 풀빼기 방법.
  20. 제1내지 8항중 어느 한 항에 따른 변이 α-아밀라제로 구성되는 것을 특징으로하는 전분 분해 조성물.
  21. 제1내지 8항중 어느 한 항에 따른 변이 α-아밀라제로 구성되는 것을 특징으로 하는 직물 풀빼기 조성물.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019910700227A 1989-06-29 1990-06-27 온도, 산 및/또는 알칼리에 대한 안정성이 증가된 변이체 미생물 알파-아밀라아제 KR0165550B1 (ko)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP89201735.1 1989-06-29
EP89201735 1989-06-29
PCT/EP1990/001042 WO1991000353A2 (en) 1989-06-29 1990-06-27 MUTANT MICROBIAL α-AMYLASES WITH INCREASED THERMAL, ACID AND/OR ALKALINE STABILITY

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KR920701449A true KR920701449A (ko) 1992-08-11
KR0165550B1 KR0165550B1 (ko) 1999-01-15

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AU638263B2 (en) 1993-06-24
DD301620A9 (de) 1993-04-29
EP0410498B1 (en) 1998-06-03
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WO1991000353A3 (en) 1991-02-21
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CN1050220A (zh) 1991-03-27
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BR9006818A (pt) 1991-08-06
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AU5953890A (en) 1991-01-17
BG61081B1 (bg) 1996-10-31
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PT94560B (pt) 1998-01-30
PT94560A (pt) 1991-02-08
EP0410498A2 (en) 1991-01-30
US5364782A (en) 1994-11-15
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