Water → (2S)-2-(carbamoylamino)-4-(methylsulfanyl)butanoic acid

Participants

Reaction Steps

Mixtures of (1) 2.0 ml. of an aqueous substrate solution prepared by dissolving DL-5-(2-methylthioethyl)hydantoin in a 0.1 M NH4Cl-NH4OH buffer solution of pH 9.5 (substrate concentration: 2.0%) and (2) 2.0 ml. of the above cell suspension were prepared and placed in test tubes, respectively. The hydrolysis reaction was then carried out at 30° C. for one hour without shaking. Immediately after the completion of the reaction, 1.0 ml. of a 10% aqueous solution of trichloroacetic acid, 1.0 ml. of a 10% solution of p-dimethylaminobenzaldehyde in 6 N hydrochloric acid and 6.0 ml. of distilled water were added to each reaction mixture. Each resulting color-developed reaction mixture was centrifuged to remove insoluble materials, and the amounts of N-carbamoylmethionine produced in the reaction mixtures were colorimetrically determined by measuring the absorbance at 420 nm. As a Control, the above procedures were repeated except that no hydantoin compound was employed.

Provenance & Evidence