Acetic Acid → L-Phenylalanine

Participants

Reaction Steps

Ten microliters of 0.2 M piperazine-N,N'-bis(2-ethanesulfonic acid) [=PIPES] buffer, pH 7.3, and 10 μl of 100 mM N-chloroacetyl-L-phenylalanine (adjusted to pH 6-7 with NaOH) are dissolved in 20 μl of water. After 10 μl of an enzyme solution (30-50 μg/ml) is added, the mixture is incubated at 37° C. for 15 minutes. Then, 50 μl of 50% acetic acid is added to stop the enzymatic reaction. The amount of L-phenylalanine produced from the substrate in the reaction solution is determined by the Yemm-Cocking ninhydrin colorimetric method (E. W. Yemm & E. C. Cocking: Analyst, Vol. 80, page 209, 1955) or with an amino acid automatic analyzer (Model 835-50, Hitachi Limited).

Provenance & Evidence