Engineering Escherichia coli for l-homoserine production.
Escherichia coli
hok/sok system
l-homoserine
metabolic engineering
plasmid stability
Journal
Journal of basic microbiology
ISSN: 1521-4028
Titre abrégé: J Basic Microbiol
Pays: Germany
ID NLM: 8503885
Informations de publication
Date de publication:
Feb 2023
Feb 2023
Historique:
revised:
13
09
2022
received:
14
08
2022
accepted:
01
10
2022
pubmed:
27
10
2022
medline:
3
2
2023
entrez:
26
10
2022
Statut:
ppublish
Résumé
l-homoserine, a nonprotein amino acid, is used to synthesize many active substances in the industry. Here, to develop a robust l-homoserine-producing strain, Escherichia coli W3110 was used as a chassis to be engineered. Based on a previous construct with blocked competing routes for l-homoserine synthesis, five genes were overexpressed by promoter replacement strategy to increase the l-homoserine production, including enhancement of precursors for l-homoserine synthesis (ppc, thrA, and asd), reinforcement of the NADPH supply (pntAB) and efflux transporters (rhtA) to improve the l-homoserine production. However, the plasmid losing was to blame for the wildly fluctuating fermentation performance of engineered strains, ranging between 2.1 and 6.2 g/L. Then, a hok/sok toxin/antitoxin system was introduced into the free plasmid expression cassette to maintain the genetic stability of the episomal plasmid; consequently, the plasmid-losing rate sharply decreased, resulting in the engineered strain SHL17, which exhibited excellent stability in l-homoserine production, with 6.3 g/L in shake flasks and 44.4 g/L in a 5-L fermenter without antibiotic addition. This work verified the effective use of the hok/sok toxin/antitoxin system combined with promoter engineering to improve the genetic stability of E. coli episomal plasmids without antibiotics.
Identifiants
pubmed: 36284486
doi: 10.1002/jobm.202200488
doi:
Substances chimiques
Homoserine
6KA95X0IVO
Escherichia coli Proteins
0
Anti-Bacterial Agents
0
Antitoxins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
168-178Subventions
Organisme : National Natural Science Foundation of China
ID : 31500043
Organisme : National Key Research and Development Program of China
ID : 2021YFC2100300
Organisme : Natural Science Foundation of Shanghai
ID : 21ZR1417200
Organisme : Open Funding Project of the State Key Laboratory of Bioreactor Engineering
Informations de copyright
© 2022 Wiley-VCH GmbH.
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