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FI105695B - Analoger av subtilisin, förfarande för framställning av dessa, deras användning och DNA-sekvens som kodar för dessa - Google Patents

Analoger av subtilisin, förfarande för framställning av dessa, deras användning och DNA-sekvens som kodar för dessa Download PDF

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Publication number
FI105695B
FI105695B FI885719A FI885719A FI105695B FI 105695 B FI105695 B FI 105695B FI 885719 A FI885719 A FI 885719A FI 885719 A FI885719 A FI 885719A FI 105695 B FI105695 B FI 105695B
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Prior art keywords
subtilisin
asn
ser
analog
asp
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FI885719A
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English (en)
Finnish (fi)
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FI885719A0 (fi
FI885719A (fi
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Yitzhak Stabinsky
Mark M Zukowski
Michael Levitt
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Amgen Inc
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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
    • 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)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
    • 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
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Cell Biology (AREA)
  • Detergent Compositions (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Cosmetics (AREA)

Claims (22)

1.- Subtilisinanalog, kännetecknad av att den har en aminosyrasekvens av det i naturen förekommande 5 Bacillus-subtilisinet, vilken modifierats genom att: (1) ersätta en eller flera av aminosyrorna Asp41, Leu75, Asn7*, Asn77, Ser7B, Ile79, Gly80, Val81, Thr208 och Tyr214, vilka är närvarande vid ett kalciumbindningsställe av det i naturen förekommande Bacillus-subtilisinet, med ίο aminosyran Asp eller Glu; och (2) avlägsna eller ersätta den ena eller bägge Asn- och Gly-aminosyrorna i vilken som heist Asn-Gly- sekvens av det i _naturen förekommande Bacillus- subtilisinet med en oiik,neutral aiifatisk aminosyra.
2. Subtilisinanalog enligt patentkrav 1, kän netecknad av att __analogen är en analog av ett i naturen förekommande_____Bacillus-subtilisin, vilken valts frän en grupp bildad av subtilisin Carlsberg, subtilisin DY, subtilisin BPN' och ett aprA-subtilisin av Bacillus 20 subtilis. _________
3. Subtilisinanalog enligt patentkrav 1, kän netecknad av att Asn76 ersatts med Asp76.
4. Subtilisinanalog enligt patentkrav 1, kännetecknad av att Asn77 ersatts med Asp77.
5. Subtilisinanalog enligt patentkrav 1, kän netecknad av att Ile79 ersatts med Glu79.
6. Subtilisinanalog enligt patentkrav 1, kän netecknad av att Asn7* ersatts med Asp7* och Asn77 med Asp77.
7. Subtilisinanalog enligt patentkrav 1, kän netecknad av att Asn76 ersatts med Asp76 och Ile79 med Glu79.
8. Subtilisinanalog enligt patentkrav 1, kän netecknad av att en Asn-rest i Asn-Gly-sekvensen er- 35 satts med en rest av en olik aminosyra. • · 105695
9. Subtilisinanalog enligt patentkrav 8, kän- netecknad av att Asn-resten i Asn-Gly-sekvensen er- satts med en Ser-rest.
10. Subtilisinanalog enligt patentkrav 9, kän- 5 netecknad av att Asn-resten i ställning 109 ersatts med en Ser-rest.
11. Subtilisinanalog enligt patentkrav 9, kän- netecknad av att Asn-resten i ställning 218 ersatts med en Ser-rest. io
12. Subtilisinanalog enligt patentkrav 9, kän- netecknad av att Asn-resten i ställningarna 109 och 218 ersatts med en Ser-rest.
13. Subtilisinanalog enligt patentkrav 12, kän-netecknad av att den valts frän en grupp bildad av 15 [Asp76, Ser109, Ser218]-subtilisin, [Asp77, Ser109, Ser218]- subtilisin, [Glu79, Ser109, Ser218]-subtil isin, [Asp76, Asp77, Ser109, Ser218]-subtilisin och [Asp76, Glu79, Ser109, Ser218]- subtilisin.
14. Subtilisinanalog enligt patentkrav 1, kän - 20 netecknad av att det modifierbara Bacillus-sub- tilisinet har en i naturen förekommande aminosyrasekvens som visas i tabell 1.
15. Subtilisinanalog enligt patentkrav 14, [Asp76, Ser109, Ser218]-subtilisin.
16. Subtilisinanalog enligt patentkrav 14, [Asp77, Ser109, Ser218]-subtilisin.
17. Subtilisinanalog enligt patentkrav 14, [Glu79, Ser109, Ser218] - subtil isin.
18. Subtilisinanalog enligt patentkrav 14, [Asp76,
30 Asp77, Ser109, Ser218]-subtilisin.
19. Subtilisinanalog enligt patentkrav 14, [Asp76, Glu79, Ser109, Ser218]-subtilisin.
20. DNA-sekvens, kännetecknad av att den kodar en subtilisinanalog med en aminosyrasekvens av det i 35 naturen förekommande Bacillus-subtilisinet, vilken sekvens modifierats genom att: • · 51 105695 (1) ersätta en eller flera av aminosyrorna Asp41, Leu75, Asn76, Asn77, Ser78, Ile79, Gly80, Vai81, Thr208 och Tyr214, vilka är närvarande vid ett kalciumbindningsställe av det i naturen förekommande Bacillus-subtilisinet, med 5 aminosyran Asp eller Glu; och (2) avlägsna eller ersätta den ena eller bägge Asn- och Gly-aminosyrorna i vilken som heist Asn-Gly-sekvens av det i naturen förekommande Bacillus-sub-tilisinet med en oiik neutral alifatisk aminosyra. ίο
21. Förfarande för att framställa Bacillus-sub- tilisin med förbättrad värme- och pH-stabilitet och omf attande ett kalciumbindningsställe, kännetecknat av att en DNA-sekvens enligt patentkrav 20 införs i en bakterie-värdcell, där den bringas att expressera och 15 nämnda Bacillus-subtilisin tillvaratas.
22. Användning av subtilisinanalog enligt patentkrav 1 i en tvättmedelssammansättning. * • «
FI885719A 1987-04-10 1988-12-09 Analoger av subtilisin, förfarande för framställning av dessa, deras användning och DNA-sekvens som kodar för dessa FI105695B (sv)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US3687287 1987-04-10
US07/036,872 US4914031A (en) 1987-04-10 1987-04-10 Subtilisin analogs
PCT/US1988/001038 WO1988008033A1 (en) 1987-04-10 1988-03-28 Subtilisin analogs
US8801038 1988-03-28

Publications (3)

Publication Number Publication Date
FI885719A0 FI885719A0 (fi) 1988-12-09
FI885719A FI885719A (fi) 1988-12-09
FI105695B true FI105695B (sv) 2000-09-29

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FI885719A FI105695B (sv) 1987-04-10 1988-12-09 Analoger av subtilisin, förfarande för framställning av dessa, deras användning och DNA-sekvens som kodar för dessa

Country Status (15)

Country Link
US (1) US4914031A (sv)
EP (1) EP0309565B2 (sv)
JP (1) JP2590247B2 (sv)
KR (1) KR960006120B1 (sv)
AT (1) ATE119941T1 (sv)
AU (1) AU612653B2 (sv)
CA (1) CA1316473C (sv)
DE (1) DE3853328T3 (sv)
DK (1) DK175576B1 (sv)
FI (1) FI105695B (sv)
HK (1) HK1007168A1 (sv)
IL (1) IL85953A (sv)
LV (1) LV11043B (sv)
NO (1) NO179679C (sv)
WO (1) WO1988008033A1 (sv)

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EP0309565A1 (en) 1989-04-05
DE3853328T3 (de) 2002-06-06
IL85953A (en) 1993-01-31
DE3853328T2 (de) 1995-07-27
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DK686788D0 (da) 1988-12-09
NO885484L (no) 1989-02-09
IL85953A0 (en) 1988-09-30
KR890700668A (ko) 1989-04-26
DK175576B1 (da) 2004-12-13
KR960006120B1 (ko) 1996-05-09
EP0309565B2 (en) 2001-08-22
DE3853328D1 (de) 1995-04-20
DK686788A (da) 1988-12-09
EP0309565B1 (en) 1995-03-15
LV11043A (lv) 1996-02-20
NO179679B (no) 1996-08-19
US4914031A (en) 1990-04-03
CA1316473C (en) 1993-04-20
LV11043B (en) 1996-06-20
JP2590247B2 (ja) 1997-03-12
FI885719A (fi) 1988-12-09
HK1007168A1 (en) 1999-04-01
EP0309565A4 (sv) 1990-02-05
JPH01502957A (ja) 1989-10-12
NO179679C (no) 1996-11-27
NO885484D0 (no) 1988-12-09
AU612653B2 (en) 1991-07-18
ATE119941T1 (de) 1995-04-15
AU1701688A (en) 1988-11-04
WO1988008033A1 (en) 1988-10-20

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