Development of highly efficient electrocompetent cells for electroporation of Geobacillus thermoglucosidasius NCIMB 11955 / 生物工程学报
Chinese Journal of Biotechnology
; (12): 3508-3519, 2023.
Article
en Zh
| WPRIM
| ID: wpr-1007973
Biblioteca responsable:
WPRO
ABSTRACT
Geobacillus thermoglucosidasius is a kind of Gram-positive facultative anaerobic bacteria. The fast growth rate under high temperature and less susceptibility to microbial contamination enable G. thermoglucosidasius to be a desirable producer of biofuels and high-value-added chemicals for the next-generation industrial biotechnology. However, compared with the classical model strain Escherichia coli, the applications of G. thermoglucosidasius are hampered by its low transformation efficiency. This study aimed at obtaining competent cells with high transformation efficiency through inactivating restriction enzymes, adding cell membrane inhibitors and cell wall weakening agents. The results showed that the electro-transformation efficiency achieved 1.2×104 CFU/(μg DNA) by knocking out four genes encoding restriction enzymes. Adding a certain amount of tween 80, dl-threonine and glycine further increased the competent efficiency about 22.5, 44, and 334 times, respectively. The electro-transformation efficiency was enhanced to 4.6×106 CFU/(μg DNA) under the optimized conditions, laying a foundation for genetic manipulation and metabolic engineering of G. thermoglucosidasius.
Palabras clave
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Bacillaceae
/
Membrana Celular
/
Electroporación
/
Escherichia coli
/
Terapia de Electroporación
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2023
Tipo del documento:
Article