THE CYANASE OPERON AND CYANATE METABOLISM

Cyanase is an inducible enzyme in C. coli that catalyzed bicarbonate-dependent decomposition of cyanate. It is encoded as part of an operon we have name the cyn operon, which includes three genes in the following order: cynT (cyanate permease), cynS (cyanase), and cynX (protein of unknown function). The direction of transcription is opposite to that of the lac operon, and the 3'-end of the cyn operon overlaps the 3'-end of the lac operon by 98 nucleotides. The gene cynR (regulatory protein) is located upstream from the cyn operon, and its transcription is opposite that of the cyn operon. The genes of the cyn operon and the cynR gene have been cloned, sequenced and over-expressed. Cyanate at concentrations of about 1 mM is toxic to strains of E. coli lacking the cyanase gene, but strains in which the inducible gene for cyanase is present can grow on cyanate as the sole source of nitrogen at concentrations as high as 20 mM. The presence of cyanase itself is not sufficient to overcome cyanate toxicity - the permease must also be present. Strains lacking the cyanase gene by having a functional permease gene, are extremely sensitive to cyanate. Uptake of cyanate involved the product of the permease gene in an energy-dependent process. It appears the the cyn operon has evolved to function in detoxification/decomposition of cyanate arising from both intra- and extracellular sources.