Phosphate → 2-Nitrophenol

Participants

Reaction Steps

That is, 0.1 ml of the present enzyme solution is added to 2.4 ml of 0.1M phosphate buffer (pH 6.5) containing 1.50% o-nitrophenyl-β-D-galactopyranoside (hereinafter referred to as "ONPG") and reacted at 70° C. for 10 minutes. Thereafter, the reaction is discontinued by the addition of 2.5 ml of 10% Na2CO3 solution. The amount of o-nitrophenol formed is obtained from the absorbance at 420 nm, and the amount of enzyme liberating 1 μmol of o-nitrophenol per minute is taken as one unit.

Provenance & Evidence