Título: | Plasmid DNA production in proteome-reduced Escherichia coli |
Autor(es): | DE LA CRUZ HERNANDEZ, MITZI RAMIREZ CAMPOS, ELISA ALEJANDRA SIGALA ALANIS, JUAN CARLOS UTRILLA CARRERI, JOSE LARA RODRIGUEZ, ALVARO RAUL |
Temas: | Celdas mínimas Reducción de proteoma Proteoma magro ADN plasmídico |
Fecha: | 2020 |
Editorial: | Suiza : MDPI |
Citation: | Microorganisms, 8, (9) 1444, 2020 |
Resumen: | The design of optimal cell factories requires engineering resource allocation for maximizing product synthesis. A recently developed method to maximize the saving in cell resources released 0.5% of the proteome of Escherichia coli by deleting only three transcription factors. We assessed the capacity for plasmid DNA (pDNA) production in the proteome-reduced strain in a mineral medium, lysogeny, and terrific broths. In all three cases, the pDNA yield from biomass was between 33 and 53% higher in the proteome-reduced than in its wild type strain. When cultured in fed-batch mode in shake-flask, the proteome-reduced strain produced 74.8 mg L−1 pDNA, which was four times greater than its wild-type strain. Nevertheless, the pDNA supercoiled fraction was less than 60% in all cases. Deletion of recA increased the pDNA yields in the wild type, but not in the proteome-reduced strain. Furthermore, recA mutants produced a higher fraction of supercoiled pDNA, compared to their parents. These results show that the novel proteome reduction approach is a promising starting point for the design of improved pDNA production hosts. |
URI: | http://ilitia.cua.uam.mx:8080/jspui/handle/123456789/769 |
Aparece en las colecciones: | Artículos |
Fichero | Descripción | Tamaño | Formato | |
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Plasmid DNA Production in Proteome-Reduced Escherichia coli.pdf | 5.72 MB | Adobe PDF | Visualizar/Abrir |
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