Title:
CORYNEBACTERIUM GLUTAMICUM EFFICIENT MUTANT, RECOMBINANT BACTERIUM CONSTRUCTION METHOD AND USE
Document Type and Number:
WIPO Patent Application WO/2024/000364
Kind Code:
A1
Abstract:
The present invention relates to the technical field of gene engineering and mutagenesis, and provides a corynebacterium glutamicum (C. glutamicum) efficient mutant, a recombinant bacterium construction method and a use. A highly conserved Exo I region is obtained on the basis of screening of a DNA polymerase from a self-DNA replication process of C. glutamicum, and by means of site-directed mutagenesis, the proofreading and repairing functions of the DNA polymerase are disabled, so that the mutation rate is improved. On the basis of a DNA polymerase mutant obtained by means of screening, a uracil DNA glycosylase inhibitor (UGI), a cytosine deaminase pmCDA1, and an adenine deaminase TadA-ABE8e are gradually connected in series and assembled, so as to construct a series of mutant plasmids (MPs). The mutant achieves an efficient mutation rate and a wide mutational spectrum in C. glutamicum, the mutation process is green and healthy, and no toxic or harmful substance participates in the mutation process. A C. glutamicum efficient mutation system constructed in the present invention can be used for screening high-yield glutamic acid host cells, and also provides an important reference for use in the production of other amino acids.
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Inventors:
RAO ZHIMING (CN)
LI XIANGFEI (CN)
QIAO ZHINA (CN)
WANG QING (CN)
XU MEIJUAN (CN)
YANG TAOWEI (CN)
ZHANG XIAN (CN)
LI XIANGFEI (CN)
QIAO ZHINA (CN)
WANG QING (CN)
XU MEIJUAN (CN)
YANG TAOWEI (CN)
ZHANG XIAN (CN)
Application Number:
PCT/CN2022/102703
Publication Date:
January 04, 2024
Filing Date:
June 30, 2022
Export Citation:
Assignee:
UNIV JIANGNAN (CN)
International Classes:
C12N1/21; C12N15/33; C12N15/54; C12N15/55; C12N15/77; C12P13/14; C12R1/15
Domestic Patent References:
| WO2021217597A1 | 2021-11-04 |
Foreign References:
| CN111411092A | 2020-07-14 | |||
| CN101698853A | 2010-04-28 | |||
| US7332310B2 | 2008-02-19 |
Other References:
DATABASE Protein 25 May 2020 (2020-05-25), ANONYMOUS : "DNA polymerase III subunit epsilon [Corynebacterium glutamicum]", XP093121953, retrieved from NCBI Database accession no. WP_011014261.1
I. J. FIJALKOWSKA ET AL.: "Mutants in the Exo I motif of Escherichia coli dnaQ: defective proofreading and inviability due to error catastrophe", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, USA, vol. 93, no. 7, 2 April 1996 (1996-04-02), XP002173814, ISSN: 0027-8424, DOI: 10.1073/pnas.93.7.2856
ZHAI HAOTIAN, QI QINGSHENG, HOU JIN: "Recent advances of continuous in vivo evolution", CHINESE JOURNAL OF BIOTECHNOLOGY, vol. 37, no. 2, 25 February 2021 (2021-02-25), pages 486 - 499, XP093121958, ISSN: 1872-2075, DOI: 10.13345/j.cjb.200311
I. J. FIJALKOWSKA ET AL.: "Mutants in the Exo I motif of Escherichia coli dnaQ: defective proofreading and inviability due to error catastrophe", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, USA, vol. 93, no. 7, 2 April 1996 (1996-04-02), XP002173814, ISSN: 0027-8424, DOI: 10.1073/pnas.93.7.2856
ZHAI HAOTIAN, QI QINGSHENG, HOU JIN: "Recent advances of continuous in vivo evolution", CHINESE JOURNAL OF BIOTECHNOLOGY, vol. 37, no. 2, 25 February 2021 (2021-02-25), pages 486 - 499, XP093121958, ISSN: 1872-2075, DOI: 10.13345/j.cjb.200311
Attorney, Agent or Firm:
BEIJING GAOWO LAW FIRM (CN)
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