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KMID : 0545120090190111408
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Journal of Microbiology and Biotechnology
2009 Volume.19 No. 11 p.1408 ~ p.1414
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Influence of Growth Conditions on Plasmid DNA Production
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Silva Filomena
Passarinha L. A.
Fani Sousa
J. A. Queiroz
Fernanda C. Domingues
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Abstract
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The obtention of high yields of purified plasmid DNA is viewed as an essential issue to be considered towards efficient production of DNA vaccines and therapeutic plasmids. In this work, Escherichia coli DH5￥a bearing the pVAX1-LacZ plasmid was grown in a developed semi-defined medium at different temperatures and tryptone concentrations. Analysis of pDNA yields and E. coli morphology revealed that at higher temperatures (37 and 40oC), higher specific yields and E. coli filamentation were obtained. However, the best results were achieved when a lower tryptone concentration was used. This approach was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure, and favoring the attainment of quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved and a specific yield of 20.43 mg pDNA/g dcw was achieved, proving that this strategy can improve pDNA yield even at a small scale.
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KEYWORD
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Plasmid DNA, fermentation, filamentous bacteria, microbial growth, temperature, tryptone
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