Bicarbonate is a recycling substrate for cyanase

Cyanase catalyzes HCO/sub 3//sup -/-dependent decomposition of cyanate to NH/sub 4//sup +/ and HCO/sub 3//sup -/. Previous studies provided evidence that carbamate is an initial product and that the kinetic mechanism is rapid equilibrium random (HCO/sub 3//sup -/ serving as substrate as opposed to activator). Direct evidence for this mechanism was obtained in this study by (1) identifying the products formed from (/sup 14/C)HCO/sub 3//sup -/ or (/sup 14/C)OCN/sup -/, (2) identifying the products formed from (/sup 13/C)HCO/sub 3//sup -/ in the presence of (/sup 18/O)H/sub 2/O, and (3) determining whether CO/sub 2/ or HCO/sub 3//sup -/ serve as substrate and are formed as product. Carbon dioxide (not HCO/sub 3//sup -/) is produced in stoichiometric amounts from both HCO/sub 3//sup -/ and OCN/sup -/. Bicarbonate (not CO/sub 2/) is the substrate. Oxygen-18 from (/sup 18/O)H/sub 2/O is not incorporated into CO/sub 2/ formed from either HCO/sub 3//sup -/ or OCN/sup -/. These results indicate that decomposition of cyanate is not a hydrolysis reaction and that HCO/sub 3//sup -/ functions as a substrate.