US20010034045A1 - Increased production of secreted proteins by recombinant eukaryotic cells - Google Patents

Increased production of secreted proteins by recombinant eukaryotic cells Download PDF

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Publication number
US20010034045A1
US20010034045A1 US09/816,277 US81627701A US2001034045A1 US 20010034045 A1 US20010034045 A1 US 20010034045A1 US 81627701 A US81627701 A US 81627701A US 2001034045 A1 US2001034045 A1 US 2001034045A1
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Prior art keywords
protein
nucleic acid
hac1
acid sequence
set forth
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Abandoned
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US09/816,277
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Merja Penttila
Michael Ward
Huaming Wang
Mari Valkonen
Markku Saloheimo
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Danisco US Inc
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Genencor International Inc
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Priority to US09/816,277 priority Critical patent/US20010034045A1/en
Assigned to GENENCOR INTERNATIONAL, INC. reassignment GENENCOR INTERNATIONAL, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: PENTTILA, MERJA E., SALOHEIMO, MARKKU L.A., VALKONEN, MARI J., WANG, HUAMING, WARD, MICHAEL
Publication of US20010034045A1 publication Critical patent/US20010034045A1/en
Priority to US10/663,450 priority patent/US7662584B2/en
Priority to US12/639,921 priority patent/US8753866B2/en
Priority to US14/272,629 priority patent/US20140335591A1/en
Abandoned legal-status Critical Current

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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
    • 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)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/11—Protein-serine/threonine kinases (2.7.11)
    • C12Y207/11001—Non-specific serine/threonine protein kinase (2.7.11.1), i.e. casein kinase or checkpoint kinase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03016—Phosphoprotein phosphatase (3.1.3.16), i.e. calcineurin

Definitions

  • FIG. 20 depicts a map of the plasmid pMS136, where the T. reesei HAC1 cDNA without the 5′ flanking region and the 20 bp intron is under the A. nidulans gpdA promoter in the vector pAN52-Notl.
  • FIG. 31 The levels of laccase activity measured in supernatants from duplicate cultures of strain ⁇ AP4:pGPTlaccase (ctrl) and transformants (#1, #2, #3, #4, #5, #6, #7 and #8) of this strain with pMS152.
  • the cell in which the proteins are secreted is any cell having an upregulated protein response.
  • the host to be transformed with the genes of the invention can be any eukaryotic cell suitable for foreign or endogenous protein production, e.g., any S. cerevisiae yeast strain, (e.g., DBY746, BMA64-1A, AH22, S150-2B, GYPY55-15bA, vtt-a-63015) any Trichoderma spp. such as T. longibrachiatum and the T.
  • bipUPREll containing the UPREll of the bip1 promoter, the element is in a reverse orientation in the promoter (unpublished results) 5′ GGG CGT GGA GAA GCG AGA AGT GGC CTC TTC TTC TCC 3′ (SEQ ID No. 45)
  • the results (FIG. 16) show that the HACl-malE fusion protein binds to the putative UPR element area found from the pdi1 promoter whereas the malE protein alone does not show any binding.
  • the binding of the fusion protein is specific, since it is competed by an excess of unlabelled oligonucleotide.
  • the fusion protein binds specifically also to the oligonucleotide pdiUPREll and not at all to pdiUPREl, and this indicates that the functional UPR element of the pdi1 promoter is UPREll.
  • the HACl-malE fusion protein also binds specifically to both of the putative UPR elements found in the bip1 promoter. Alignment of the three T. reesei UPR element shows that the consensus sequence for binding is GC(C/G)A(G/A)N 1-2 GTG(G/T)C (FIG. 16) (SEQ ID No. 46).

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Mycology (AREA)
  • Biophysics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
US09/816,277 2000-03-24 2001-03-23 Increased production of secreted proteins by recombinant eukaryotic cells Abandoned US20010034045A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
US09/816,277 US20010034045A1 (en) 2000-03-24 2001-03-23 Increased production of secreted proteins by recombinant eukaryotic cells
US10/663,450 US7662584B2 (en) 2000-03-24 2003-09-15 Increased production of secreted proteins by recombinant eukaryotic cells
US12/639,921 US8753866B2 (en) 2000-03-24 2009-12-16 Increased production of secreted proteins by recombinant eukaryotic cells
US14/272,629 US20140335591A1 (en) 2000-03-24 2014-05-08 Increased production of secreted proteins by recombinant eukaryotic cells

Applications Claiming Priority (2)

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US53469200A 2000-03-24 2000-03-24
US09/816,277 US20010034045A1 (en) 2000-03-24 2001-03-23 Increased production of secreted proteins by recombinant eukaryotic cells

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US53469200A Continuation-In-Part 2000-03-24 2000-03-24

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US09/816,277 Abandoned US20010034045A1 (en) 2000-03-24 2001-03-23 Increased production of secreted proteins by recombinant eukaryotic cells
US10/663,450 Expired - Fee Related US7662584B2 (en) 2000-03-24 2003-09-15 Increased production of secreted proteins by recombinant eukaryotic cells
US12/639,921 Expired - Fee Related US8753866B2 (en) 2000-03-24 2009-12-16 Increased production of secreted proteins by recombinant eukaryotic cells
US14/272,629 Abandoned US20140335591A1 (en) 2000-03-24 2014-05-08 Increased production of secreted proteins by recombinant eukaryotic cells

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US10/663,450 Expired - Fee Related US7662584B2 (en) 2000-03-24 2003-09-15 Increased production of secreted proteins by recombinant eukaryotic cells
US12/639,921 Expired - Fee Related US8753866B2 (en) 2000-03-24 2009-12-16 Increased production of secreted proteins by recombinant eukaryotic cells
US14/272,629 Abandoned US20140335591A1 (en) 2000-03-24 2014-05-08 Increased production of secreted proteins by recombinant eukaryotic cells

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US (4) US20010034045A1 (de)
EP (1) EP1266018B1 (de)
AT (1) ATE394496T1 (de)
AU (1) AU2001250954A1 (de)
CA (1) CA2403842C (de)
DE (1) DE60133884D1 (de)
DK (1) DK1266018T3 (de)
WO (1) WO2001072783A2 (de)

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US6607904B2 (en) 1998-09-30 2003-08-19 Danisco A/S Peptidyl prolyl cis-trans isomerases
US20060053502A1 (en) * 2004-09-02 2006-03-09 Yijie Gao Systems and methods for protein production
US20080193977A1 (en) * 2003-12-23 2008-08-14 Novozymes Delta Limited Gene Expression Technique
US7479379B2 (en) 2004-03-31 2009-01-20 Centocor, Inc. Methods for altering protein production rates
US20100093030A1 (en) * 2006-11-02 2010-04-15 Cornelis Maria Jacobus Sagt Production of secreted proteins by filamentous fungi
WO2010122141A1 (en) 2009-04-24 2010-10-28 Dsm Ip Assets B.V. Carbohydrate degrading polypeptide and uses thereof
WO2010121933A1 (en) 2009-04-22 2010-10-28 Dsm Ip Assets B.V. Process for the production of a recombinant polypeptide of interest
WO2011009700A1 (en) 2009-07-22 2011-01-27 Dsm Ip Assets B.V. Improved host cell for the production of a compound of interest
WO2011095374A2 (en) 2010-02-04 2011-08-11 Dsm Ip Assets B.V. Process for preparing filamentous fungal strains having a sexual cycle and a process for preparing sexually crossed filamentous fungal strains
WO2011098580A1 (en) 2010-02-11 2011-08-18 Dsm Ip Assets B.V. Polypeptide having cellobiohydrolase activity and uses thereof
WO2011098577A1 (en) 2010-02-11 2011-08-18 Dsm Ip Assets B.V. Host cell capable of producing enzymes useful for degradation of lignocellulosic material
WO2012000886A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having beta-glucosidase activity and uses thereof
WO2012000888A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having acetyl xylan esterase activity and uses thereof
WO2012001169A1 (en) 2010-07-01 2012-01-05 Dsm Ip Assets B.V. A method for the production of a compound of interest
WO2012000891A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having carbohydrate degrading activity and uses thereof
WO2012000890A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having beta-glucosidase activity and uses thereof
WO2012000887A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having swollenin activity and uses thereof
WO2012000892A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having or assisting in carbohydrate material degrading activity and uses thereof
EP2410048A1 (de) 2005-01-24 2012-01-25 DSM IP Assets B.V. Verfahren zur Herstellung einer Verbindung in einer filamentösen Pilzzelle
EP2423315A1 (de) 2006-06-29 2012-02-29 DSM IP Assets B.V. Verfahren für verbesserte Polypeptidexpression
EP2458007A1 (de) 2005-11-29 2012-05-30 DSM IP Assets B.V. DNA-Bindungsstelle eines transkriptionellen Aktivators zur Genexpression
EP2631295A2 (de) 2007-02-15 2013-08-28 DSM IP Assets B.V. Rekombinante Hostzelle zur Herstellung einer Verbindung von Interesse
US20130252286A1 (en) * 2010-11-05 2013-09-26 Asahi Glass Company, Limited Transformant of yeast of genus schizosaccharomyces, and method for producing same
WO2014013074A1 (en) 2012-07-19 2014-01-23 Dsm Ip Assets B.V. Agse-deficient strain
WO2018166943A1 (en) 2017-03-13 2018-09-20 Dsm Ip Assets B.V. Zinc binuclear cluster transcriptional regulator-deficient strain
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JP5131666B2 (ja) * 2006-05-16 2013-01-30 独立行政法人産業技術総合研究所 タンパク質の高分泌生産方法
KR100977446B1 (ko) * 2006-06-16 2010-08-24 한국생명공학연구원 분비 스트레스 반응을 조절하는 한세눌라 폴리모르파의신규한 유전자 및 상기 유전자를 이용하여 재조합 단백질의분비 발현 효율을 증가시키는 방법
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US8546126B2 (en) * 2007-03-21 2013-10-01 Danisco Us Inc. Over expression of foldases and chaperones improves protein production
CA2704548A1 (en) * 2007-11-01 2009-05-07 Danisco Us Inc. Signal sequences and co-expressed chaperones for improving protein production in a host cell
CN101952303A (zh) * 2008-02-25 2011-01-19 诺维信公司 提高目标蛋白质的表达产率的方法
EP2258854A1 (de) 2009-05-20 2010-12-08 FH Campus Wien Eukaryotische Wirtszelle mit einem Ausdrucksverstärker
FI20105633A0 (fi) 2010-06-04 2010-06-04 Valtion Teknillinen Parannettu menetelmä proteiinien tuottamiseksi
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US6607904B2 (en) 1998-09-30 2003-08-19 Danisco A/S Peptidyl prolyl cis-trans isomerases
US6638737B1 (en) * 1998-09-30 2003-10-28 Danisco A/S Peptidyl prolyl cis-trans isomerases
US20080193977A1 (en) * 2003-12-23 2008-08-14 Novozymes Delta Limited Gene Expression Technique
US8969064B2 (en) 2003-12-23 2015-03-03 Novozymes Biopharma Dk A/S Gene expression technique
US9057061B2 (en) 2003-12-23 2015-06-16 Novozymes Biopharma Dk A/S Gene expression technique
US7479379B2 (en) 2004-03-31 2009-01-20 Centocor, Inc. Methods for altering protein production rates
WO2005094355A3 (en) * 2004-03-31 2009-03-05 Centocor Inc Methods for altering protein production rates
KR101145308B1 (ko) * 2004-03-31 2012-05-14 얀센 바이오테크 인코포레이티드 단백질 생산 속도를 변경하는 방법
AU2005228444B2 (en) * 2004-03-31 2010-11-25 Centocor, Inc. Methods for altering protein production rates
EP2339015A1 (de) * 2004-03-31 2011-06-29 Centocor Ortho Biotech Inc. Verfahren zur Änderung der Proteinproduktionsraten
US7514237B2 (en) 2004-09-02 2009-04-07 Wyeth Systems and methods for protein production
US20090221035A1 (en) * 2004-09-02 2009-09-03 Wyeth Systems and methods for protein production
US20060053502A1 (en) * 2004-09-02 2006-03-09 Yijie Gao Systems and methods for protein production
US8173394B2 (en) 2004-09-02 2012-05-08 Wyeth Llc Systems and methods for protein production
EP2410048A1 (de) 2005-01-24 2012-01-25 DSM IP Assets B.V. Verfahren zur Herstellung einer Verbindung in einer filamentösen Pilzzelle
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EP2458007A1 (de) 2005-11-29 2012-05-30 DSM IP Assets B.V. DNA-Bindungsstelle eines transkriptionellen Aktivators zur Genexpression
EP2423315A1 (de) 2006-06-29 2012-02-29 DSM IP Assets B.V. Verfahren für verbesserte Polypeptidexpression
US8389269B2 (en) 2006-11-02 2013-03-05 Dsm Ip Assets B.V. Production of secreted proteins by filamentous fungi
US20100093030A1 (en) * 2006-11-02 2010-04-15 Cornelis Maria Jacobus Sagt Production of secreted proteins by filamentous fungi
EP2631295A2 (de) 2007-02-15 2013-08-28 DSM IP Assets B.V. Rekombinante Hostzelle zur Herstellung einer Verbindung von Interesse
WO2010121933A1 (en) 2009-04-22 2010-10-28 Dsm Ip Assets B.V. Process for the production of a recombinant polypeptide of interest
WO2010122141A1 (en) 2009-04-24 2010-10-28 Dsm Ip Assets B.V. Carbohydrate degrading polypeptide and uses thereof
EP3293264A1 (de) 2009-07-22 2018-03-14 DSM IP Assets B.V. Verbesserte wirtszelle für die herstellung von relevanten produkten
WO2011009700A1 (en) 2009-07-22 2011-01-27 Dsm Ip Assets B.V. Improved host cell for the production of a compound of interest
WO2011095374A2 (en) 2010-02-04 2011-08-11 Dsm Ip Assets B.V. Process for preparing filamentous fungal strains having a sexual cycle and a process for preparing sexually crossed filamentous fungal strains
WO2011098580A1 (en) 2010-02-11 2011-08-18 Dsm Ip Assets B.V. Polypeptide having cellobiohydrolase activity and uses thereof
WO2011098577A1 (en) 2010-02-11 2011-08-18 Dsm Ip Assets B.V. Host cell capable of producing enzymes useful for degradation of lignocellulosic material
WO2012000888A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having acetyl xylan esterase activity and uses thereof
WO2012000886A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having beta-glucosidase activity and uses thereof
WO2012000892A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having or assisting in carbohydrate material degrading activity and uses thereof
WO2012000891A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having carbohydrate degrading activity and uses thereof
WO2012000890A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having beta-glucosidase activity and uses thereof
WO2012000887A1 (en) 2010-06-29 2012-01-05 Dsm Ip Assets B.V. Polypeptide having swollenin activity and uses thereof
WO2012001169A1 (en) 2010-07-01 2012-01-05 Dsm Ip Assets B.V. A method for the production of a compound of interest
US20130252286A1 (en) * 2010-11-05 2013-09-26 Asahi Glass Company, Limited Transformant of yeast of genus schizosaccharomyces, and method for producing same
WO2014013074A1 (en) 2012-07-19 2014-01-23 Dsm Ip Assets B.V. Agse-deficient strain
WO2018166943A1 (en) 2017-03-13 2018-09-20 Dsm Ip Assets B.V. Zinc binuclear cluster transcriptional regulator-deficient strain
CN113166211A (zh) * 2018-11-28 2021-07-23 诺维信公司 改善的丝状真菌宿主细胞

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AU2001250954A1 (en) 2001-10-08
EP1266018A2 (de) 2002-12-18
US8753866B2 (en) 2014-06-17
US20140335591A1 (en) 2014-11-13
CA2403842C (en) 2012-09-25
US20100221775A1 (en) 2010-09-02
DE60133884D1 (de) 2008-06-19
WO2001072783A2 (en) 2001-10-04
CA2403842A1 (en) 2001-10-04
EP1266018B1 (de) 2008-05-07
US20040186070A1 (en) 2004-09-23

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