Títol:
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Development of a highly efficient production process for recombinant protein expression in Escherichia coli NEB10β
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Autor/a:
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Miret, Joan; Román, Ramón; Benito Peinado, Mario; Casablancas, Antoni; Guillén, Marina; Álvaro, Gregorio; González Anadón, Gloria
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Abstract:
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Altres ajuts: COST Action CM 1303 - Systems Biocatalysis |
Abstract:
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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. |
Matèries:
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-E. coli NEB10β -PTDH-CHMO -High cell culture -PBAD expression system -Fed-Batch |
Drets:
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open access
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https://creativecommons.org/licenses/by-nc-nd/4.0/ |
Tipus de document:
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Article |
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Uri:
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https://ddd.uab.cat/record/261062
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