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KMID : 1094720070120030213
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Biotechnology and Bioprocess Engineering
2007 Volume.12 No. 3 p.213 ~ p.221
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Optimization of medium components for plasmid production by recombinant E. coli DH5α pUK21CMVβ1.2
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Zheng Shu-Zhen
Friehs Karl
He Ning
Deng Xu
Li Qing-Biao
He Zheng
Xu Chen-Yun
Lu Ying-Hua
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Abstract
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In this study we investigated the effects of carbon and nitrogen sources, and the carbon to nitrogen ratio (C:N) of growth medium on the production of plasmid DNA by pUK21CMVβ1.2 harbouring recombinantE. coli. The results indicated that sucrose and casein peptone were the most suitable carbon and nitrogen sources, respectively, for plasmid production, and the plasmid concentration was significantly influenced by the C:N ratio. A Box-Behnken design and surface response methodology were employed to formulate the optimal medium. In the optimized medium, a plasmid concentration of 51.80 mg/L and a plasmid selectivity of 23.49 mg/g were achieved after 24 h of incubation. These values were approximately 3.5 and 1.7 times higher, respectively, than those obtained on the original LB medium.
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KEYWORD
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plasmid DNA, medium optimization, single-factor experiments, response surface methodology, statistical analysis
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