Title:
RECOMBINANT ESCHERICHIA COLI FOR EFFICIENT PRODUCTION OF GLUTARIC ACID, AND CONSTRUCTION METHOD THEREFOR AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/092632
Kind Code:
A1
Abstract:
Provided are a recombinant Escherichia coli for efficient production of glutaric acid, and a construction method therefor and the use thereof. A two-plasmid recombinant bacterium is constructed by means of molecular biology; an aromatic aldehyde synthase (AAS) gene, an amine oxidase (Mao) gene and an aldehyde dehydrogenase (Glox) gene are co-expressed; and the constructed expression plasmids are introduced into Escherichia coli to construct a recombinant cell. Then, screening is performed by means of an ampicillin-resistant and kanamycin-resistant combined plate, and the recombinant strain for the efficient production of glutaric acid is obtained. By optimizing the substrate concentration, bacterial concentration and transformation temperature, the efficient production of glutaric acid is achieved. Under the transformation conditions where the bacterium has a concentration of 30 gL and a pH of 8 and 6 mM of NAD+ is additionally added, reaction is performed for 30 h, 30 gL of L-lysine can be transformed into 19.65 g of glutaric acid, and the transformation rate can reach 65.3%.
Inventors:
LIU LIMING (CN)
ZHANG ZHILAN (CN)
GAO CONG (CN)
CHEN XIULAI (CN)
GUO LIANG (CN)
LIU JIA (CN)
SONG WEI (CN)
ZHANG ZHILAN (CN)
GAO CONG (CN)
CHEN XIULAI (CN)
GUO LIANG (CN)
LIU JIA (CN)
SONG WEI (CN)
Application Number:
PCT/CN2021/135262
Publication Date:
June 01, 2023
Filing Date:
December 03, 2021
Export Citation:
Assignee:
UNIV JIANGNAN (CN)
International Classes:
C12N15/70; C12N1/21; C12P7/44; C12R1/19
Domestic Patent References:
| WO2005089505A2 | 2005-09-29 |
Foreign References:
| CN111849845A | 2020-10-30 | |||
| CN112226398A | 2021-01-15 | |||
| CN108753636A | 2018-11-06 |
Other References:
WANG, JIAPING ET AL.: "Engineering the Cad pathway in Escherichia coli to produce glutarate from L-lysine.", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY., vol. 105, no. 9, 27 April 2021 (2021-04-27), XP037445517, DOI: 10.1007/s00253-021-11275-1
ZENG JIAOJIAO, YU SHIQIN; ZHOU JINGWEN: "Metabolic Engineering of Escherichia Coli for Improving Tyrosol Production", FOOD AND FERMENTATION INDUSTRIES, vol. 47, no. 22, 28 April 2021 (2021-04-28), pages 8 - 15, XP093068959, DOI: 10.13995/j.cnki.11-1802/ts.027055
LI WENNA, MA LIN, SHEN XIAOLIN, WANG JIA, FENG QI, LIU LEXUAN, ZHENG GUOJUN, YAN YAJUN, SUN XINXIAO, YUAN QIPENG: "Targeting metabolic driving and intermediate influx in lysine catabolism for high-level glutarate production", NATURE COMMUNICATIONS, vol. 10, no. 1, XP093068962, DOI: 10.1038/s41467-019-11289-4
DATABASE PROTEIN ANONYMOUS : "aldehyde dehydrogenase family 3 member A2 isoform 2 [Homo sapiens]", XP093068964, retrieved from NCBI
WANG JIAPING, GAO CONG, CHEN XIULAI, LIU LIMING: "Expanding the lysine industry: biotechnological production of L-lysine and its derivatives", ADVANCES IN APPLIED MICROBIOLOGY, vol. 115, 18 March 2021 (2021-03-18), pages 1 - 33, XP009546666, ISSN: 0065-2164, DOI: 10.1016/bs.aambs.2021.02.001
ZENG JIAOJIAO, YU SHIQIN; ZHOU JINGWEN: "Metabolic Engineering of Escherichia Coli for Improving Tyrosol Production", FOOD AND FERMENTATION INDUSTRIES, vol. 47, no. 22, 28 April 2021 (2021-04-28), pages 8 - 15, XP093068959, DOI: 10.13995/j.cnki.11-1802/ts.027055
LI WENNA, MA LIN, SHEN XIAOLIN, WANG JIA, FENG QI, LIU LEXUAN, ZHENG GUOJUN, YAN YAJUN, SUN XINXIAO, YUAN QIPENG: "Targeting metabolic driving and intermediate influx in lysine catabolism for high-level glutarate production", NATURE COMMUNICATIONS, vol. 10, no. 1, XP093068962, DOI: 10.1038/s41467-019-11289-4
DATABASE PROTEIN ANONYMOUS : "aldehyde dehydrogenase family 3 member A2 isoform 2 [Homo sapiens]", XP093068964, retrieved from NCBI
WANG JIAPING, GAO CONG, CHEN XIULAI, LIU LIMING: "Expanding the lysine industry: biotechnological production of L-lysine and its derivatives", ADVANCES IN APPLIED MICROBIOLOGY, vol. 115, 18 March 2021 (2021-03-18), pages 1 - 33, XP009546666, ISSN: 0065-2164, DOI: 10.1016/bs.aambs.2021.02.001
Attorney, Agent or Firm:
CENTRAL SOUTH WELL INTELLECTUAL PROPERTY OFFICE (CN)
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