DK1012307T3 - Autocatalytically activatable zymogenic protease precursors and their use - Google Patents

Autocatalytically activatable zymogenic protease precursors and their use

Info

Publication number
DK1012307T3
DK1012307T3 DK98945214T DK98945214T DK1012307T3 DK 1012307 T3 DK1012307 T3 DK 1012307T3 DK 98945214 T DK98945214 T DK 98945214T DK 98945214 T DK98945214 T DK 98945214T DK 1012307 T3 DK1012307 T3 DK 1012307T3
Authority
DK
Denmark
Prior art keywords
protease
zymogenic
precursor
host cell
autocatalytically
Prior art date
Application number
DK98945214T
Other languages
Danish (da)
Inventor
Erhard Kopetzki
Wolfram Bode
Robert Huber
Karl-Peter Hopfner
Original Assignee
Roche Diagnostics Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Roche Diagnostics Gmbh filed Critical Roche Diagnostics Gmbh
Application granted granted Critical
Publication of DK1012307T3 publication Critical patent/DK1012307T3/en

Links

Classifications

    • 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)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • 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)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/647—Blood coagulation factors not provided for in a preceding group or according to more than one of the proceeding groups
    • 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)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6427—Chymotrypsins (3.4.21.1; 3.4.21.2); Trypsin (3.4.21.4)
    • 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)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6432—Coagulation factor Xa (3.4.21.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21006—Coagulation factor Xa (3.4.21.6)

Abstract

A process for the recombinant production of a protease is characterised by (a) transformation of a host cell with a recombinant nucleic acid which codes for a zymogenic protease precursor containing an autocatalytic cleavage site which does not occur naturally, this cleavage site being recognised by the active form of said protease, cleaving the zymogenic precursor into the active protease; (b) cultivation of the host cell such that the zymogenic protease precursor is created in the host cell in the form of inclusion bodies; (c) isolation and naturalisation of the inclusion bodies in such conditions that the protease part of the zymogenic precursor is produced with its natural conformation; and (d) autocatalytic cleavage of the naturalised zymogenic precursor to produce the active protease. This process is suitable to produce recombinant proteases in a simple manner and with high yields.
DK98945214T 1997-08-22 1998-08-12 Autocatalytically activatable zymogenic protease precursors and their use DK1012307T3 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP97114513 1997-08-22
EP97117816 1997-10-15
PCT/EP1998/005094 WO1999010503A1 (en) 1997-08-22 1998-08-12 Zymogenic protease precursors that can be autocatalytically activated and their use
EP98945214A EP1012307B1 (en) 1997-08-22 1998-08-12 Zymogenic protease precursors that can be autocatalytically activated and their use

Publications (1)

Publication Number Publication Date
DK1012307T3 true DK1012307T3 (en) 2005-09-05

Family

ID=26145739

Family Applications (1)

Application Number Title Priority Date Filing Date
DK98945214T DK1012307T3 (en) 1997-08-22 1998-08-12 Autocatalytically activatable zymogenic protease precursors and their use

Country Status (9)

Country Link
EP (1) EP1012307B1 (en)
JP (1) JP3947653B2 (en)
AT (1) ATE294869T1 (en)
AU (1) AU9260598A (en)
CA (1) CA2301698A1 (en)
DE (1) DE59812768D1 (en)
DK (1) DK1012307T3 (en)
IL (1) IL134653A0 (en)
WO (1) WO1999010503A1 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6159722A (en) 1997-12-03 2000-12-12 Boehringer Mannheim Gmbh Chimeric serine proteases
GB9911689D0 (en) * 1999-05-19 1999-07-21 Medical Res Council Refolding method
TR200200253T2 (en) * 1999-08-09 2002-05-21 Bio-Chemie Gesellschaft M.B.H. Production of proteins
CN1230542C (en) * 1999-08-09 2005-12-07 山德士股份公司 Production of proteins by autoproteolytic cleavage
AU7053500A (en) * 1999-09-15 2001-04-17 Eli Lilly And Company Chymotrypsin-free trypsin
WO2001066771A2 (en) * 2000-03-03 2001-09-13 Zymogenetics, Inc. Human serine protease
CA2437342C (en) 2001-02-01 2013-08-06 F. Hoffmann-La Roche Ag Method for producing recombinant trypsin
DE60328802D1 (en) 2002-08-30 2009-09-24 Novozymes Inc PROCESS FOR THE PREPARATION OF MAMMALS TRYPSINS
EP1558731A4 (en) * 2002-10-04 2007-01-10 Schering Ag Modified hepsin molecules having a substitute activation sequence and uses thereof
US7176013B2 (en) 2003-06-11 2007-02-13 Schering Aktiengesellschaft Modified corin molecules having substitute activation sequences and uses thereof
JP2015092852A (en) * 2013-11-12 2015-05-18 倉敷紡績株式会社 Composition for biological tissue dispersion
JPWO2018174231A1 (en) * 2017-03-24 2020-01-23 株式会社カネカ Method for producing protease in yeast

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA1340740C (en) * 1987-12-08 1999-09-14 Eileen R. Mulvihill Co-expression in eukaryotic cells
DE4140699A1 (en) * 1991-01-11 1992-07-16 Boehringer Mannheim Gmbh RECOMBINANT IGA PROTEASE
IL107536A0 (en) * 1992-11-13 1994-02-27 Lilly Co Eli Expression vectors for bovine trypsin and trypsinogen and host cells transformed therewith
JP3920331B2 (en) * 1994-12-09 2007-05-30 ノボ ノルディスク アクティーゼルスカブ Method for producing extracellular proteins in bacteria
AU6121796A (en) * 1995-06-16 1997-01-15 Novo Nordisk A/S A process for producing trypsin (trypsinogen)

Also Published As

Publication number Publication date
ATE294869T1 (en) 2005-05-15
EP1012307B1 (en) 2005-05-04
DE59812768D1 (en) 2005-06-09
WO1999010503A1 (en) 1999-03-04
EP1012307A1 (en) 2000-06-28
IL134653A0 (en) 2001-04-30
AU9260598A (en) 1999-03-16
JP2001514003A (en) 2001-09-11
JP3947653B2 (en) 2007-07-25
CA2301698A1 (en) 1999-03-04

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