Phosphate → p-Cyanobenzamide

Participants

Reaction Steps

The nitrile hydratase activity was measured as follows. The cells (about 1 g by wet mass) were added to a reaction solution obtained by suspending from 1 to 10 mass % of terephthalonitrile (TPN) as a substrate in 10 ml of 20 mM phosphate buffer solution (pH: 7.0) and reacted at 30° C. while shaking, and the p-cyanobenzoic acid amide produced in the reaction solution was quantitated by HPLC at fixed intervals. The solid matter was removed from the reaction solution by centrifugation, and the supernatant 100-fold diluted with the eluant was used as the HPLC sample. The amidase activity was measured by using p-cyanobenzamide or benzamide serving as a substrate, performing the reaction under the same conditions as above, and determining the generated p-cyanobenzoic acid or benzoic acid by HPLC.

Provenance & Evidence