Global Patent Index - EP 1451367 A4

EP 1451367 A4 20060614 - ENGINEERING OF LEADER PEPTIDES FOR THE SECRETION OF RECOMBINANT PROTEINS IN BACTERIA

Title (en)

ENGINEERING OF LEADER PEPTIDES FOR THE SECRETION OF RECOMBINANT PROTEINS IN BACTERIA

Title (de)

ENGINEERING VON LEADER-PEPTIDEN FÜR DIE SEKRETION VON REKOMBINANTEN PROTEINEN IN BAKTERIEN

Title (fr)

GENIE DE PEPTIDES LEADER POUR LA SECRETION DE PROTEINES RECOMBINEES DANS DES BACTERIES

Publication

EP 1451367 A4 20060614 (EN)

Application

EP 02795597 A 20021105

Priority

  • US 0235618 W 20021105
  • US 33745201 P 20011105

Abstract (en)

[origin: WO03040335A2] The present invention provides methods of isolating of leader peptides capable of directing export of heterologous proteins from the bacterial cytoplasm. The methods rely on the screening of libraries of putative leader peptides or of leader peptide mutants for sequences that allow rapid export and thus can rescue a short-lived reporter protein from degradation in the cytoplasm. The mutant leader peptides identified herein are shown to confer significantly higher steady state levels of export not only for short-lived reporter protein but also for other stable, long-lived proteins. These leader peptides can be used to direct or enhance protein secretion. The present invention further discloses methods for the export of cytoplasmically folded protein via the Tat pathway. Proteins having disulfide bonds are first folded within the cytoplasm in suitable oxidizing mutant strains. Such cytoplasmically pre-folded proteins containing disulfide bonds are then exported via the Tat pathway.

IPC 1-7

C12Q 1/68

IPC 8 full level

C07K 14/435 (2006.01); C12N 15/10 (2006.01); C12N 15/62 (2006.01); C12P 21/02 (2006.01)

CPC (source: EP)

C07K 14/43595 (2013.01); C12N 15/1051 (2013.01); C12N 15/625 (2013.01); C12P 21/02 (2013.01); C07K 2319/034 (2013.01); C07K 2319/60 (2013.01); C07K 2319/61 (2013.01); C07K 2319/95 (2013.01)

Citation (search report)

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  • [PXPY] WO 0222667 A2 20020321 - GENENCOR INT [US], et al
  • [PX] WO 02055717 A2 20020718 - GENENCOR INT [US], et al
  • [X] WO 9945136 A1 19990910 - UNIV BRITISH COLUMBIA [CA]
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  • [XY] MATTEUCCI M ET AL: "ALKALINE PHOSPHATASE FUSIONS: A TAG TO IDENTIFY MUTATIONS THAT RESULT IN INCREASED EXPRESSION OF SECRETED HUMAN GROWTH HORMONE FROM E. COLI", BIO/TECHNOLOGY, NATURE PUBLISHING CO. NEW YORK, US, vol. 4, January 1986 (1986-01-01), pages 51 - 55, XP002006627, ISSN: 0733-222X
  • [XY] SANTINI C-L ET AL: "Translocation of jellyfish green fluorescent protein via the Tat system of Escherichia coli and change of its periplasmic localization in response to osmotic up-shock", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY OF BIOLOGICAL CHEMISTS, BALTIMORE, MD, US, vol. 276, no. 11, 16 March 2001 (2001-03-16), pages 8159 - 8164, XP002273739, ISSN: 0021-9258
  • [XY] THOMAS JOANNA D ET AL: "Export of active green fluorescent protein to the periplasm by the twin-arginine translocase (Tat) pathway in Escherichia coli", MOLECULAR MICROBIOLOGY, vol. 39, no. 1, January 2001 (2001-01-01), pages 47 - 53, XP002330124, ISSN: 0950-382X
  • [XY] HINSLEY^A A P ET AL: "A naturally occurring bacterial Tat signal peptide lacking one of the 'invariant' arginine residues of the consensus targeting motif", FEBS LETTERS, ELSEVIER SCIENCE PUBLISHERS, AMSTERDAM, NL, vol. 497, no. 1, 18 May 2001 (2001-05-18), pages 45 - 49, XP004250034, ISSN: 0014-5793
  • [Y] FEILMEIER BRADLEY J ET AL: "Green fluorescent protein functions as a reporter for protein localization in Escherichia coli", JOURNAL OF BACTERIOLOGY, WASHINGTON, DC, US, vol. 182, no. 14, July 2000 (2000-07-01), pages 4068 - 4076, XP002263489, ISSN: 0021-9193
  • [XY] BUCHANAN G ET AL: "A genetic screen for suppressors of escherichia coli tat signal peptide mutations establishes a critical role for the second arginine within the twin-arginine motif", ARCHIVES OF MICROBIOLOGY, BERLIN, DE, vol. 177, 30 October 2001 (2001-10-30), pages 107 - 112, XP002971648, ISSN: 0302-8933
  • [Y] ANDERSEN ET AL: "New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, WASHINGTON,DC, US, vol. 64, no. 6, June 1998 (1998-06-01), pages 2240 - 2246, XP002203612, ISSN: 0099-2240
  • [A] BERG B L ET AL: "NITRATE-INDUCIBLE FORMATE DEHYDROGENASE IN ESCHERICHIA-COLI K-12 I. NUCLEOTIDE SEQUENCE OF THE FDN GHI OPERON AND EVIDENCE THAT OPAL UGA ENCODES SELENOCYSTEINE", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 266, no. 33, 1991, pages 22380 - 22385, XP002330126, ISSN: 0021-9258
  • [X] BLAUDECK N ET AL: "Specificity of signal peptide recognition in tat-dependent bacterial protein translocation", JOURNAL OF BACTERIOLOGY, WASHINGTON, DC, US, vol. 183, no. 2, January 2001 (2001-01-01), pages 604 - 610, XP002989193, ISSN: 0021-9193
  • [PX] IZE B ET AL: "In vivo dissection of the Tat translocation pathway in Escherichia coli", JOURNAL OF MOLECULAR BIOLOGY, LONDON, GB, vol. 317, no. 3, 29 March 2002 (2002-03-29), pages 327 - 335, XP004468971, ISSN: 0022-2836
  • [T] DELISA MATTHEW P ET AL: "Phage shock protein PspA of Escherichia coli relieves saturation of protein export via the Tat pathway.", JOURNAL OF BACTERIOLOGY, vol. 186, no. 2, January 2004 (2004-01-01), pages 366 - 373, XP002378230, ISSN: 0021-9193
  • See references of WO 03040335A2

Designated contracting state (EPC)

AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LI LU MC NL PT SE SK TR

DOCDB simple family (publication)

WO 03040335 A2 20030515; WO 03040335 A3 20031231; CN 100564540 C 20091202; CN 1788092 A 20060614; EP 1451367 A2 20040901; EP 1451367 A4 20060614

DOCDB simple family (application)

US 0235618 W 20021105; CN 02826734 A 20021105; EP 02795597 A 20021105