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KMID : 0545119920020010035
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Journal of Microbiology and Biotechnology
1992 Volume.2 No. 1 p.35 ~ p.40
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Influence of Plasmid Properties on Fermentation Parameters of Recombinant Escherichia coli
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Lee, In Young
Seo, Dong Jin/Lee, Sun Bok
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Abstract
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The influence of the nature of plasmids on fermentation parameters such as cell growth, cell viability, plasmid stability, and product formation has been investigated using E. coli M5248 and its recombinant derivatives M5248 [pBR322], M5248[pAS1], and M5248[pNKM21]. At a low temperature (30℃), the cell growth, cell viability, and protein synthesis of the recombinants were nearly identical to those of the host cell. However, at high temperature (42℃), in which transcription from the P_L promoter is derepressed, the recombinant cells showed decreased stability along with lower growth rates and cell viability. The ratio of total protein to cell mass was in the order of E. coli M5248>M5248[pBR322]>M5248[pAS1]>M5248[pNKM21]. It was found that transcription from the P_L promoter adversely affect the plasmid maintenance and host cell metabolism even in the absence of the cloned-gene expression. Furthermore, profiles of β-lactamase activity were shown to vary with recombinant strains. E coli M5248[pBR322] showed highest β-lactamase activity at 30℃, while at 42℃ β-lactamase activity was significantly reduced irrespective of the strains. The effect of the plasmid properties on plasmid-encoded gene expression has been further examined based on the relationship between β-lactamse activity and plasmid-harboring cell numbers.
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