SUMMARY: The effect of temperature on the stability of pTG201, a plasmid carrying the gene (which encodes catechol 2,3-dioxygenase from ), and the production of catechol 2,3-dioxygenase in free and immobilized during continuous culture have been studied at various temperatures. Immobilization of cells increased the stability of pTG201 considerably, even under conditions when expression of the product was enhanced. Since transcription was controlled by the γP promoter and 857 repressor, increasing derepression temperatures increased catechol 2,3-dioxygenase productivity and decreased pTG201 stability. A two-stage continuous culture system to overcome the impact of the high-level expression of the gene on the stability of pTG201 is described. In the first stage, immobilized cells were grown in the repressed state in order to prevent loss of pTG201, whereas in the second stage, cultures were maintained in the derepressed state.


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