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Pàgina inicial > Articles > Articles publicats > Development of a highly efficient production process for recombinant protein expression in Escherichia coli NEB10β |
Data: | 2020 |
Resum: | Recombinant protein expression in E. coli is well described, with multiple strains and process strategies available. However, strains used for cloning and molecular biology purposes are not generally considered for protein expression. Using these strains could result in a simplification of the production pathway of a newly cloned protein of interest. In this work, the E. coli strain NEB10β has been characterized for the expression of the complex fusion protein phosphite dehydrogenase-cyclohexanone monooxygenase (PTDH-CHMO), and a production process has been developed based on the PBAD expression system. A fed-batch approach using a defined medium supplemented with amino acids, with glycerol as a carbon source, allows for an efficient recombinant protein expression process, incrementing 9. 2-fold the production obtained in a complex medium batch and reaching around 2 g/L of product after 6 h of induction. The process was successfully reproduced in a NEB10β strain for the production of the alcohol dehydrogenase (ADH) enzyme. |
Ajuts: | European Commission 635734 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-1462 |
Nota: | Altres ajuts: COST Action CM 1303 - Systems Biocatalysis |
Drets: | Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades. |
Llengua: | Anglès |
Document: | Article ; recerca ; Versió acceptada per publicar |
Matèria: | E. coli NEB10β ; PTDH-CHMO ; High cell culture ; PBAD expression system ; Fed-Batch |
Publicat a: | Biochemical engineering journal, Vol. 159 (July 2020) , art. 107612, ISSN 1369-703X |
Postprint 19 p, 463.2 KB |