Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production of ColE1 Type Plasmid by Escherichia coli $DH5\alpha$ Cultured Under Nonselective Conditions

  • PASSARINHA L. A. (Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico, Departamento de Quimica, Universidade da Beira Interior) ;
  • DIOGO M. M. (Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico) ;
  • QUEIROZ J. A. (Departamento de Quimica, Universidade da Beira Interior) ;
  • MONTEIRO G. A. (Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico) ;
  • FONSECA L. P. (Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico) ;
  • PRAZERES D. M. F. (Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico)
  • Published : 2006.01.01
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Abstract

Plasmid DNA (pDNA) is a product of interest for many biopharmaceutical companies and research laboratories, because of increase in the number of gene therapy protocols that use nonviral vectors. This work was undertaken to study the effect of antibiotic and dissolved oxygen concentration (DOC) on the production of a ColE 1-type plasmid (pVAX1-LacZ) hosted in Escherichia coli $DH5\alpha$ and cultured in a batch fermentor with 0.751 of Terrific Broth. A decrease in the DOC from $60\%\;to\;5\%$ was shown to increase the specific pDNA concentration approximately 1.5-fold, due to the downregulation of growth. Additionally, this increase in the pDNA concentration led to a 2.2-fold increase in the purity of cell lysates obtained after cell lysis. However, the use of higher DOC led to 2.8-fold higher volumetric productivity as a consequence of a faster growth rate, reducing the fermentation time from 24 to 8 h. Interestingly, the specific pDNA concentration, and pDNA productivity and purity were always higher $(10-15\%)$ in the absence of antibiotic. Overall, the data indicate that nonselective conditions can be used without compromising yield, productivity, and purity of pDNA.

Keywords

References

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