WO2009126463A3 - Filamentous fungi with inactivated protease genes for altered protein production - Google Patents

Filamentous fungi with inactivated protease genes for altered protein production Download PDF

Info

Publication number
WO2009126463A3
WO2009126463A3 PCT/US2009/038626 US2009038626W WO2009126463A3 WO 2009126463 A3 WO2009126463 A3 WO 2009126463A3 US 2009038626 W US2009038626 W US 2009038626W WO 2009126463 A3 WO2009126463 A3 WO 2009126463A3
Authority
WO
WIPO (PCT)
Prior art keywords
protein production
filamentous fungi
altered protein
protease genes
inactivated protease
Prior art date
Application number
PCT/US2009/038626
Other languages
French (fr)
Other versions
WO2009126463A2 (en
Inventor
Huaming Wang
Original Assignee
Danisco Us Inc., Genencor Division
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 Danisco Us Inc., Genencor Division filed Critical Danisco Us Inc., Genencor Division
Priority to CN2009801122959A priority Critical patent/CN102099460A/en
Priority to EP09729753A priority patent/EP2262888A2/en
Priority to CA2720827A priority patent/CA2720827A1/en
Priority to JP2011504054A priority patent/JP2011517943A/en
Publication of WO2009126463A2 publication Critical patent/WO2009126463A2/en
Publication of WO2009126463A3 publication Critical patent/WO2009126463A3/en

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • C07K14/38Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from Aspergillus
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/48Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/58Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi
    • C12N9/62Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi from Aspergillus
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • C12P21/02Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione

Abstract

The invention relates to a filamentous fungal cell (e.g., Aspergillus sp.) comprising at least one inactivated protease gene chosen from apsB, a homolog of apsB, cpsA, a homolog cpsA, and combinations thereof. Nucleic acids and methods for making the inactivated mutant filamentous fungal cells are provided as well as methods for using the cells for the altered production of endogenous or heterologous proteins of interest.
PCT/US2009/038626 2008-04-08 2009-03-27 Filamentous fungi with inactivated protease genes for altered protein production WO2009126463A2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
CN2009801122959A CN102099460A (en) 2008-04-08 2009-03-27 Filamentous fungi with inactivated protease genes for altered protein production
EP09729753A EP2262888A2 (en) 2008-04-08 2009-03-27 Filamentous fungi with inactivated protease genes for altered protein production
CA2720827A CA2720827A1 (en) 2008-04-08 2009-03-27 Filamentous fungi with inactivated protease genes for altered protein production
JP2011504054A JP2011517943A (en) 2008-04-08 2009-03-27 Filamentous fungi with inactivated protease genes for the production of modified proteins

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US4328408P 2008-04-08 2008-04-08
US61/043,284 2008-04-08

Publications (2)

Publication Number Publication Date
WO2009126463A2 WO2009126463A2 (en) 2009-10-15
WO2009126463A3 true WO2009126463A3 (en) 2010-01-14

Family

ID=40848716

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2009/038626 WO2009126463A2 (en) 2008-04-08 2009-03-27 Filamentous fungi with inactivated protease genes for altered protein production

Country Status (6)

Country Link
US (2) US20090253173A1 (en)
EP (1) EP2262888A2 (en)
JP (1) JP2011517943A (en)
CN (1) CN102099460A (en)
CA (1) CA2720827A1 (en)
WO (1) WO2009126463A2 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012048334A2 (en) * 2010-10-08 2012-04-12 Dyadic International (Usa) Inc. Novel fungal proteases
EP2760997A4 (en) 2011-09-30 2015-02-11 Codexis Inc Fungal proteases
BR112014016587A2 (en) 2012-01-05 2020-10-27 Novartis Ag protease-deficient filamentous fungal cells and methods of using them
WO2013174927A1 (en) 2012-05-23 2013-11-28 Novartis International Pharmaceutical Limited Production of fucosylated glycoproteins
WO2013181647A2 (en) 2012-06-01 2013-12-05 Danisco Us Inc. Compositions and methods of producing isoprene and/or industrrial bio-products using anaerobic microorganisms
CA2916594C (en) * 2013-07-04 2020-03-10 Novartis Ag O-mannosyltransferase deficient filamentous fungal cells and methods of use thereof
WO2015004241A2 (en) 2013-07-10 2015-01-15 Novartis Ag Multiple proteases deficient filamentous fungal cells and methods of use thereof
WO2015054507A1 (en) * 2013-10-10 2015-04-16 Pronutria, Inc. Nutritive polypeptide production systems, and methods of manufacture and use thereof
EP3172333B1 (en) 2014-07-21 2020-05-13 Glykos Finland Oy Production of glycoproteins with mammalian-like n-glycans in filamentous fungi
CN104846090B (en) * 2015-05-12 2018-08-17 海南大学 Genetic of fungi screening technique based on marker gene
KR102593668B1 (en) * 2016-10-04 2023-10-24 다니스코 유에스 인크. Protein production in filamentous fungal cells in the absence of inducing substrates
CN110564625B (en) * 2019-08-13 2023-01-24 内蒙古世洪农业科技有限公司 Saline-alkali resistant aspergillus flavus and separation method and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997022705A1 (en) * 1995-12-15 1997-06-26 Novo Nordisk A/S A FUNGUNS WHEREIN THE areA, pepC AND/OR pepE GENES HAVE BEEN INACTIVATED
US5840570A (en) * 1988-07-01 1998-11-24 Berka; Randy M. Aspartic proteinase deficient filamentous fungi
WO1999002705A1 (en) * 1997-07-05 1999-01-21 Societe Des Produits Nestle S.A. CLONING OF THE PROLYL-DIPEPTIDYL-PEPTIDASE FROM $i(ASPERGILLUS ORYZAE)
WO2006110677A2 (en) * 2005-04-12 2006-10-19 Genencor International, Inc. Gene inactivated mutants with altered protein production
WO2007062936A2 (en) * 2005-11-29 2007-06-07 Dsm Ip Assets B.V. Dna binding site of a transcriptional activator useful in gene expression

Family Cites Families (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4760025A (en) * 1984-05-29 1988-07-26 Genencor, Inc. Modified enzymes and methods for making same
US4683202A (en) * 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4965188A (en) * 1986-08-22 1990-10-23 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme
US4683195A (en) * 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
DE69133612D1 (en) * 1990-12-10 2009-04-02 Genencor Int Improved cellulose saccharification by cloning and amplification of the Trichoderma beta-glucosidase gene
US5674728A (en) * 1993-11-03 1997-10-07 Novartis Corporation Aspergillus niger vacuolar aspartyl protease
US5861271A (en) * 1993-12-17 1999-01-19 Fowler; Timothy Cellulase enzymes and systems for their expressions
US5594119A (en) * 1994-09-20 1997-01-14 Novo Nordisk A/S Gene encoding carboxypeptidase of aspergillus niger
DE69631127T2 (en) * 1995-05-18 2004-07-08 Genencor International, Inc., Palo Alto GLYCOSYL TRANSFERASE EXPRESSION IN ASPERGILLUS
JPH11501217A (en) * 1995-08-03 1999-02-02 ギスト ブロカデス ベスローテン フェンノートシャップ Use of the homologous amdS gene as a selectable marker
US6187577B1 (en) * 1997-11-19 2001-02-13 Genecor International, Inc. Cellulase producing Actinomycetes cellulase produced therefrom and method of producing same
US6268328B1 (en) * 1998-12-18 2001-07-31 Genencor International, Inc. Variant EGIII-like cellulase compositions
AU2001234443A1 (en) * 2000-01-13 2001-07-24 Novozymes Biotech, Inc. Methods for producing a polypeptide using a crippled translational initiator sequence
US7323327B2 (en) * 2001-02-23 2008-01-29 Dsm Ip Assets B.V. Genes encoding novel proteolytic enzymes
US20040014955A1 (en) * 2001-12-17 2004-01-22 Carlos Zamudio Identification of essential genes of cryptococcus neoformans and methods of use
AU2002358179A1 (en) * 2001-12-21 2003-07-09 Dsm Ip Assets B.V. New rennets
AU2003263163A1 (en) * 2002-09-20 2004-04-08 Cym1P A/S Methods for increasing the production of a recombinant polypeptide from a host cell
JP4050712B2 (en) * 2004-02-27 2008-02-20 独立行政法人科学技術振興機構 Method for homologous recombination
JP2008507294A (en) * 2004-07-26 2008-03-13 ダウ グローバル テクノロジーズ インコーポレイティド Method for improved protein expression by strain genetic manipulation
US20060211089A1 (en) * 2005-03-16 2006-09-21 Novozymes A/S Expression of defensins in filamentous fungi
EP1930428B1 (en) * 2005-08-03 2010-12-01 Asahi Glass Company, Limited transformed yeast host cells AND METHOD OF PRODUCING FOREIGN PROTEIN
US8216844B2 (en) * 2006-08-29 2012-07-10 Danisco Us Inc. Compositions and methods for improved protein production
JP4998873B2 (en) * 2006-08-30 2012-08-15 独立行政法人酒類総合研究所 How to make filamentous fungi that produce spores with low viability

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5840570A (en) * 1988-07-01 1998-11-24 Berka; Randy M. Aspartic proteinase deficient filamentous fungi
US6509171B1 (en) * 1988-07-01 2003-01-21 Genencor International, Inc. Aspartic proteinase deficient filamentous fungi
WO1997022705A1 (en) * 1995-12-15 1997-06-26 Novo Nordisk A/S A FUNGUNS WHEREIN THE areA, pepC AND/OR pepE GENES HAVE BEEN INACTIVATED
WO1999002705A1 (en) * 1997-07-05 1999-01-21 Societe Des Produits Nestle S.A. CLONING OF THE PROLYL-DIPEPTIDYL-PEPTIDASE FROM $i(ASPERGILLUS ORYZAE)
WO2006110677A2 (en) * 2005-04-12 2006-10-19 Genencor International, Inc. Gene inactivated mutants with altered protein production
WO2007062936A2 (en) * 2005-11-29 2007-06-07 Dsm Ip Assets B.V. Dna binding site of a transcriptional activator useful in gene expression

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BARTKEVICIUTE D ET AL: "Disruption of the MNN10 gene enhances protein secretion in Kluyveromyces lactis and Saccharomyces cerevisiae", FEMS YEAST RESEARCH, ELSEVIER SCIENCE, TOKYO, NL, vol. 4, no. 8, 1 September 2004 (2004-09-01), pages 833 - 840, XP004533627, ISSN: 1567-1356 *
HOMBERGH VAN DEN J P T W ET AL: "DISRUPTION OF THREE ACID PROTEASES IN ASPERGILLUS NIGER EFFECTS ON PROTEASE SPECTRUM, INTRACELLULAR PROTEOLYSIS, AND DEGRADATION OF TARGET PROTEINS", EUROPEAN JOURNAL OF BIOCHEMISTRY, BLACKWELL PUBLISHING, BERLIN, DE, vol. 247, no. 2, 1 January 1997 (1997-01-01), pages 605 - 613, XP001153668, ISSN: 0014-2956 *
OKA TAKUJI ET AL: "Molecular characterization of protein O-mannosyltransferase and its involvement in cell-wall synthesis in Aspergillus nidulans", MICROBIOLOGY, SOCIETY FOR GENERAL MICROBIOLOGY, READING, GB, vol. 150, no. Part 6, 1 June 2004 (2004-06-01), pages 1973 - 1982, XP002399299, ISSN: 1350-0872 *
SUELMANN RUEDIGER ET AL: "Increased nuclear traffic chaos in hyphae of Aspergillus nidulans: Molecular characterization of apsB and in vivo observation of nuclear behaviour", MOLECULAR MICROBIOLOGY, vol. 30, no. 4, November 1998 (1998-11-01), pages 831 - 842, XP002537754, ISSN: 0950-382X *
VAN DEN HOMBERGH J P T W ET AL: "Aspergillus as a host for heterologous protein production: the problem of proteases", TRENDS IN BIOTECHNOLOGY, ELSEVIER PUBLICATIONS, CAMBRIDGE, GB, vol. 15, no. 7, 1 July 1997 (1997-07-01), pages 256 - 263, XP004092682, ISSN: 0167-7799 *
WANG ET AL: "Isolation of four pepsin-like protease genes from Aspergillus niger and analysis of the effect of disruptions on heterologous laccase expression", FUNGAL GENETICS AND BIOLOGY, SAN DIEGO, CA, US, vol. 45, no. 1, 15 November 2007 (2007-11-15), pages 17 - 27, XP022346803, ISSN: 1087-1845 *

Also Published As

Publication number Publication date
EP2262888A2 (en) 2010-12-22
US20090253173A1 (en) 2009-10-08
US20130266984A1 (en) 2013-10-10
CN102099460A (en) 2011-06-15
CA2720827A1 (en) 2009-10-15
JP2011517943A (en) 2011-06-23
WO2009126463A2 (en) 2009-10-15

Similar Documents

Publication Publication Date Title
WO2009126463A3 (en) Filamentous fungi with inactivated protease genes for altered protein production
WO2014205192A3 (en) Targeted integration
WO2007140339A8 (en) Production of isoprenoids
CA3033327A1 (en) Programmable cas9-recombinase fusion proteins and uses thereof
WO2008153029A1 (en) Method for expression of specific gene
WO2010039889A3 (en) Methods for using positively and negatively selectable genes in a filamentous fungal cell
WO2006073727A3 (en) Recombinant dna constructs and methods for controlling gene expression
WO2012058494A3 (en) Isoprene synthase variants for improved production of isoprene
WO2009042070A3 (en) Production of isoprenoids
WO2008104890A3 (en) Compositions and methods for producing apolipoprotein
WO2008154695A8 (en) Method for improving stress resistance in plants and materials therefor
WO2005090552A3 (en) Method of engineering a cytidine monophosphate-sialic acid synthetic pathway in fungi and yeast
WO2007146706A3 (en) Modified dicamba monooxygenase enzyme and methods of its use
WO2010107303A3 (en) Chrysosporium lucknowense protein production system
WO2008153903A3 (en) Heterologous and homologous cellulase expression system
WO2006110677A3 (en) Gene inactivated mutants with altered protein production
WO2012009272A3 (en) Pentose fermentation by a recombinant microorganism
WO2010135836A8 (en) Novel beta-glucosidase enzymes
WO2008071726A3 (en) Pathogen inducible plant trehalose-6-phophate phophatase gene promoters and regulatory elements
WO2009158627A3 (en) Cellulosic protein expression in yeast
WO2008065200A8 (en) Dnase expression in recombinant host cells
WO2012168493A3 (en) Baculoviral dna elements for the expression of recombinant proteins in a host insect
WO2011151512A3 (en) Method for protein production in filamentous fungi
MX2010009071A (en) Isolated peptides having phospholipase inhibitory activity.
WO2010127219A3 (en) Altering enzyme balance through fermentation conditions

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 200980112295.9

Country of ref document: CN

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09729753

Country of ref document: EP

Kind code of ref document: A2

WWE Wipo information: entry into national phase

Ref document number: 6303/DELNP/2010

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2009729753

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2720827

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2011504054

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE