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KMID : 0545119920020010026
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Journal of Microbiology and Biotechnology
1992 Volume.2 No. 1 p.26 ~ p.34
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Effect of Induction Temperature on the P_L Promoter Controlled Production of Recombinant Human Interleukin-2 in Escherichia coli
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Lee, In Young
Kim, Myung Kuk/Lee, Sun Bok
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Abstract
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The effect of induction temperature on fermentation parameters has been investigated extensively using Escherichia coli M5248[pNKM21], a producer of recombinant human interleukin-2 (rhIL-2). In this recombinant microorganism, the gene expression of rhIL-2 is regulated by the c1857 repressor and P_L promoter system. The recombinant fermentation parameters studied in this work include the cell growth, protein synthesis, cell viability, plasmid stability, β-lactamase activity, and rhIL-2 productivity. Interrelationships of such fermentation parameters have been analyzed through a quantitative assessment of the experimental data set obtained at eight different culture conditions. While the expression of rhIL-2 gene was repressed at culture temperatures below 34℃ with little effect on other fermentation parameters, under the conditions of rhIL-2 production (36∼44℃) the cell growth, plasmid stability, and β-lactamase activity were, as induction temperature was increased, more profoundly reduced. Although the rhIL-2 content in the insoluble protein fraction was maximum at 40℃, total rhIL-2 production in the culture volume was found to be highest at the induction temperature of 36℃. This was in contrast to the previously known optimum induction temperature of the P_L promoter system (40∼42℃). Explanations for such a discrepancy have been proposed based on a product formation kinetics, and their implications have been discussed in detail.
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KEYWORD
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