WO1991000920A3 - Process for preparing a protein by a fungus transformed by multicopy integration of an expression vector - Google Patents

Process for preparing a protein by a fungus transformed by multicopy integration of an expression vector

Info

Publication number
WO1991000920A3
WO1991000920A3 PCT/EP1990/001138 EP9001138W WO9100920A3 WO 1991000920 A3 WO1991000920 A3 WO 1991000920A3 EP 9001138 W EP9001138 W EP 9001138W WO 9100920 A3 WO9100920 A3 WO 9100920A3
Authority
WO
WIPO (PCT)
Prior art keywords
protein
expression vector
preparing
multicopy integration
integration
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/EP1990/001138
Other languages
French (fr)
Other versions
WO1991000920A2 (en
Inventor
Marco Luigi Frederic Giuseppin
Maria Teresa Santos Lopes
Roelf Johannes Planta
Johannes Maria Antoni Verbakel
Cornelis Theodorus Verrips
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Unilever NV
Original Assignee
Unilever NV
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
Priority claimed from GB898915659A external-priority patent/GB8915659D0/en
Priority to DE69031710T priority Critical patent/DE69031710T2/en
Priority to EP90911878A priority patent/EP0481008B1/en
Priority to CA002063592A priority patent/CA2063592C/en
Application filed by Unilever NV filed Critical Unilever NV
Publication of WO1991000920A2 publication Critical patent/WO1991000920A2/en
Publication of WO1991000920A3 publication Critical patent/WO1991000920A3/en
Priority to EP19910201627 priority patent/EP0464922A1/en
Priority to CA 2046249 priority patent/CA2046249A1/en
Priority to JP3261485A priority patent/JPH06292573A/en
Anticipated expiration legal-status Critical
Priority to US08/034,650 priority patent/US5641671A/en
Ceased legal-status Critical Current

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)
    • C12N9/24—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)
    • C12N9/2465—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
    • 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)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C12N9/20—Triglyceride splitting, e.g. by means of lipase

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Abstract

A process is disclosed for preparing a protein by a eukaryote transformed by multicopy integration of an expression vector into the genome of a yeast, such as Saccharomyces, Hansenula, and Kluyveromyces, or of a mould such as Aspergillus, Rhizopus and Trichoderma, said expression vector containing both an ''expressible gene'' encoding said protein and a so-called ''deficient selection marker needed for the growth of the yeast or mould in a specific medium'', such as the LEU2d, TRP1d or URA3d gene, in combination with a ribosomal DNA sequence, resulting in stable high copy integration of 100-300 copies per cell. This multicopy integration results in an increased production of the desired protein, which can be guar α-galactosidase, an oxidase or a hydrolytic enzyme such as a lipase.
PCT/EP1990/001138 1989-07-07 1990-07-09 Process for preparing a protein by a fungus transformed by multicopy integration of an expression vector Ceased WO1991000920A2 (en)

Priority Applications (7)

Application Number Priority Date Filing Date Title
DE69031710T DE69031710T2 (en) 1989-07-07 1990-07-09 METHOD FOR PRODUCING A PROTEIN BY MEANS OF A MUSHROOM TRANSFORMED BY MULTIPLE COPY INTEGRATION OF AN EXPRESSION VECTOR
EP90911878A EP0481008B1 (en) 1989-07-07 1990-07-09 Process for preparing a protein by a fungus transformed by multicopy integration of an expression vector
CA002063592A CA2063592C (en) 1989-07-07 1990-07-09 Process for preparing a protein by a fungus transformed by multicopy integration of an expression vector
EP19910201627 EP0464922A1 (en) 1990-07-06 1991-06-26 Production of active pseudomonas glumae lipase in homologous or heterologous hosts
CA 2046249 CA2046249A1 (en) 1990-07-06 1991-07-04 Production of active pseudomonas glumae lipase in homologous or heterologous hosts
JP3261485A JPH06292573A (en) 1990-07-06 1991-07-05 Production of active Pseudomonas glumae lipase in homologous or heterologous hosts
US08/034,650 US5641671A (en) 1990-07-06 1993-03-22 Production of active Pseudomonas glumae lipase in homologous or heterologous hosts

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GB8915659.0 1989-07-07
GB898915659A GB8915659D0 (en) 1989-07-07 1989-07-07 Enzymes,their production and use
EP90201007.3 1990-04-20
EP90201007 1990-04-20

Publications (2)

Publication Number Publication Date
WO1991000920A2 WO1991000920A2 (en) 1991-01-24
WO1991000920A3 true WO1991000920A3 (en) 1991-03-07

Family

ID=26125849

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1990/001138 Ceased WO1991000920A2 (en) 1989-07-07 1990-07-09 Process for preparing a protein by a fungus transformed by multicopy integration of an expression vector

Country Status (10)

Country Link
US (1) US6090574A (en)
EP (2) EP0481008B1 (en)
JP (1) JP3273609B2 (en)
AT (2) ATE196505T1 (en)
CA (1) CA2063592C (en)
DE (2) DE69031710T2 (en)
DK (2) DK0778348T3 (en)
ES (2) ES2109238T3 (en)
IE (1) IE81016B1 (en)
WO (1) WO1991000920A2 (en)

Cited By (6)

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Publication number Priority date Publication date Assignee Title
US7371423B2 (en) 1997-04-09 2008-05-13 Danisco, A/S Method for preparing flour doughs and products made from such doughs using lipase
US7622290B2 (en) 2004-03-12 2009-11-24 Danisco A/S Fungal lipolytic enzymes, nucleic acids encoding, and uses thereof
US7638293B2 (en) 2003-01-17 2009-12-29 Danisco A/S Method
USRE43135E1 (en) 2001-05-18 2012-01-24 Danisco A/S Method of improving dough and bread quality
USRE43341E1 (en) 1995-06-07 2012-05-01 Danisco A/S Method of improving the properties of a flour dough, a flour dough improving composition and improved food products
US8889371B2 (en) 2004-07-16 2014-11-18 Dupont Nutrition Biosciences Aps Lipolytic enzyme: uses thereof in the food industry

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WO1991014781A1 (en) * 1990-03-19 1991-10-03 Henkel Research Corporation METHOD FOR INCREASING THE OMEGA-HYDROXYLASE ACTIVITY IN $i(CANDIDA TROPICALIS)
ATE140974T1 (en) * 1990-10-04 1996-08-15 Japan Maize Prod HOST VECTOR SYSTEM
JP3133774B2 (en) * 1990-10-04 2001-02-13 日本食品化工株式会社 Novel host-vector system
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AU5811194A (en) * 1992-12-11 1994-07-04 Quest International B.V. The use of the (kluyveromyces marxianus) inulinase gene promoter for protein production
US5786212A (en) * 1993-09-02 1998-07-28 Pfizer Inc. Multiple integrative vectors and Yarrowia lipolytica transformants
JP2001501089A (en) * 1996-09-24 2001-01-30 バハマイア，アンドレアス DNA constructs and methods for producing proteins using these DNA constructs
US7883872B2 (en) 1996-10-10 2011-02-08 Dyadic International (Usa), Inc. Construction of highly efficient cellulase compositions for enzymatic hydrolysis of cellulose
JP4307563B2 (en) 1997-04-07 2009-08-05 ユニリーバー・ナームローゼ・ベンノートシャープ Agrobacterium-mediated transformation of filamentous fungi, especially those belonging to the genus Aspergillus
EP0870835A1 (en) * 1997-04-07 1998-10-14 Unilever N.V. Agrobacterium mediated transformation of moulds, in particular those belonging to the genus Aspergillus
JP5303084B2 (en) 1997-04-11 2013-10-02 コニンクリーケ デーエスエム ナムローゼ フェンノートシャップ Gene conversion as a means to build industrial recombinant organisms
ATE398181T1 (en) * 1997-04-11 2008-07-15 Dsm Ip Assets Bv YEAST CELLS WHICH CONTAIN AT LEAST TWO COPIES OF A DESIRED GENE AND INTEGRATE IN THE CHROMOSOMAL GENOME ON MORE THAN ONE NON-RIBOSOMAL RNA CODING AREA, ESPECIALLY WITH KLUYVEROMYCES
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ATE196505T1 (en) 2000-10-15
DE69033633D1 (en) 2000-10-26
WO1991000920A2 (en) 1991-01-24
IE81016B1 (en) 1999-09-22
ES2109238T3 (en) 1998-01-16
IE902496A1 (en) 1991-02-13
DE69031710T2 (en) 1998-03-05
EP0481008B1 (en) 1997-11-12
DK0481008T3 (en) 1998-05-04
JP3273609B2 (en) 2002-04-08
US6090574A (en) 2000-07-18
CA2063592A1 (en) 1991-01-08
EP0778348B1 (en) 2000-09-20
DE69033633T2 (en) 2001-05-03

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