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Biotechnology and Bioengineering, Vol.43, No.11, 1190-1193, 1994
Application of the Cross-Regulation System as a Metabolic Switch
The ability to switch metabolic flow from one pathway to another at a desired point in a bioprocess expands the horizons of metabolic engineering. Such an externally inducible switch can be realized by embedding synthetic operons behind two cross-regulated promoters. This results in coordinated cessation of transcription of one operon while transcription of a second operon is simultaneously activated. The ability to effect such coordinated and inverse control of transcription of two operons has been illustrated experimentally using a model construct containing two different reporter genes, Vitreoscilla hemoglobin (VHb) and chloramphenicol acetyltransferase (CAT), fused to lambda P-L and tac promoters, respectively, along with corresponding repressor genes in a cross-regulation configuration. Only VHb production was observed preinduction, and postinduction only CAT was produced. The framework presented here and its obvious extensions can be used with different combinations of promoter systems and synthetic operon constructs to achieve complicated metabolic flux regulation in diverse hosts.
Keywords:ESCHERICHIA-COLI;BREVIBACTERIUM-LACTOFERMENTUM;HOMOSERINE DEHYDROGENASE;VITREOSCILLA HEMOGLOBIN;THREONINE PRODUCTION;EXPRESSION;GENE