Improved cryptic plasmids in probiotic Escherichia coli Nissle 1917 for antibiotic-free pathway engineering.
Appl Microbiol Biotechnol
; 107(16): 5257-5267, 2023 Aug.
Article
en En
| MEDLINE
| ID: mdl-37405431
ABSTRACT
The engineered probiotic Escherichia coli Nissle 1917 (EcN) is expected to be employed in the diagnosis and treatment of various diseases. However, the introduced plasmids typically require antibiotics to maintain genetic stability, and the cryptic plasmids in EcN are usually eliminated to avoid plasmid incompatibility which may change the inherent probiotic characteristics. Here, we provided a simple design to minimize the genetic change of probiotics by eliminating native plasmids and reintroducing the recombinants carrying functional genes. Specific insertion sites in the vectors showed significant differences in the expression of fluorescence proteins. Selected integration sites were applied in the de novo synthesis of salicylic acid, leading to a titer of 142.0 ± 6.0 mg/L in a shake flask with good production stability. Additionally, the design successfully realized the biosynthesis of ergothioneine (45 mg/L) by one-step construction. This work expands the application scope of native cryptic plasmids to the easy construction of functional pathways. KEY POINTS ⢠Cryptic plasmids of EcN were designed to express exogenous genes ⢠Insertion sites with different expression intensities in cryptic plasmids were provided ⢠Target products were stably produced by engineering cryptic plasmids.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Probióticos
/
Antibacterianos
Idioma:
En
Revista:
Appl Microbiol Biotechnol
Año:
2023
Tipo del documento:
Article
País de afiliación:
China
Pais de publicación:
ALEMANHA
/
ALEMANIA
/
DE
/
DEUSTCHLAND
/
GERMANY