Title:
CRM197 PROTEIN EXPRESSION METHOD
Document Type and Number:
WIPO Patent Application WO/2021/045292
Kind Code:
A1
Abstract:
The present invention relates to a signal sequence for expressing a CRM197 protein in Escherichia coli and secreting same into the periplasm, and a use thereof, and more specifically, to: a signal sequence for expressing a CRM197 protein; a nucleic acid for coding the signal sequence; a nucleic acid construct or expression vector comprising the nucleic acid and a CRM197 protein gene; a recombinant microorganism having the nucleic acid construct or expression vector introduced therein; and a CRM197 protein production method comprising a step for culturing the recombinant microorganism. According to the present invention, a CRM197 protein having the same physicochemical/immunologic properties as the protein isolated from the parent bacteria may be expressed even in regular Escherichia coli of which a redox potential is not adjusted, and a CRM197 protein having high periplasmic secretion efficiency may be produced even without shifting the pH of a culture medium in order to increase secretion into the periplasm, and thus the present invention is very useful in CRM197 protein production.
Inventors:
KIM JEONG HYUN (KR)
KIM HYUN-DO (KR)
PARK EUN-HYE (KR)
KANG JUNG HYUN (KR)
PAN JAE GU (KR)
KIM EUI JOONG (KR)
LEE CHANKYU (KR)
PARK MINCHUL (KR)
KIM HYUN-DO (KR)
PARK EUN-HYE (KR)
KANG JUNG HYUN (KR)
PAN JAE GU (KR)
KIM EUI JOONG (KR)
LEE CHANKYU (KR)
PARK MINCHUL (KR)
Application Number:
PCT/KR2019/012899
Publication Date:
March 11, 2021
Filing Date:
October 02, 2019
Export Citation:
Assignee:
GENOFOCUS CO LTD (KR)
EUBIOLOGICS CO LTD (KR)
EUBIOLOGICS CO LTD (KR)
International Classes:
C07K7/08; C07K14/00; C07K14/34
Domestic Patent References:
| WO1991017424A1 | 1991-11-14 | |||
| WO1991016024A1 | 1991-10-31 |
Foreign References:
| KR20160128362A | 2016-11-07 | |||
| KR20030014959A | 2003-02-20 | |||
| KR20090039239A | 2009-04-22 | |||
| KR20150082968A | 2015-07-16 | |||
| US5049386A | 1991-09-17 | |||
| US4946787A | 1990-08-07 | |||
| US4897355A | 1990-01-30 | |||
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| US4485054A | 1984-11-27 | |||
| US4501728A | 1985-02-26 | |||
| US4774085A | 1988-09-27 | |||
| US4837028A | 1989-06-06 |
Other References:
CHOI, J. H. ET AL.: "Secretory and extracellular production of recombinant proteins using Escherichia coli", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 64, 14 February 2004 (2004-02-14), pages 625 - 635, XP002402621
UCHIDA ET AL., JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 248, 1973, pages 3845 - 3850
MALITO ET AL., PROC. NATL. ACAD. SCI. USA, vol. 109, no. 14, 1990, pages 2995 - 342012
BISHAI ET AL., J. BACTERIOL., vol. 169, pages 5140 - 5151
CRYSTAL, SCIENCE, vol. 270, 1995, pages 404 - 410
BLAESE ET AL., CANCER GENE THER., vol. 2, 1995, pages 291 - 297
REMY ET AL., BIOCONJUGATE CHEM., vol. 5, 1994, pages 382389 - 654
GAO ET AL., GENE THERAPY., vol. 2, 1995, pages 710 - 722
AHMAD ET AL., CANCER RES., vol. 52, 1992, pages 4817 - 4820
AMANN ET AL., GENE, vol. 69, 1988, pages 301 - 15
JEONG ET AL., APPL ENVIRON MICROBIOL., vol. 78, 2012, pages 5440 - 3
See also references of EP 4026842A4
UCHIDA ET AL., JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 248, 1973, pages 3845 - 3850
MALITO ET AL., PROC. NATL. ACAD. SCI. USA, vol. 109, no. 14, 1990, pages 2995 - 342012
BISHAI ET AL., J. BACTERIOL., vol. 169, pages 5140 - 5151
CRYSTAL, SCIENCE, vol. 270, 1995, pages 404 - 410
BLAESE ET AL., CANCER GENE THER., vol. 2, 1995, pages 291 - 297
REMY ET AL., BIOCONJUGATE CHEM., vol. 5, 1994, pages 382389 - 654
GAO ET AL., GENE THERAPY., vol. 2, 1995, pages 710 - 722
AHMAD ET AL., CANCER RES., vol. 52, 1992, pages 4817 - 4820
AMANN ET AL., GENE, vol. 69, 1988, pages 301 - 15
JEONG ET AL., APPL ENVIRON MICROBIOL., vol. 78, 2012, pages 5440 - 3
See also references of EP 4026842A4
Attorney, Agent or Firm:
LEE, Cheo Young et al. (KR)
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