Hydrochloric Acid → HCID 178508913, compound

Participants

Reaction Steps

5,10-Methylenetetrahydrofolate-dehydrogenase catalyzes the reaction, 5,10-methylene-THF+NADP+ →5,10-methenyl-THF+NADPH+H+. A 0.95 mL solution containing 1 μmole THF, 10 μmoles formaldehyde (fresh), 0.6 μmoles NADP and 25 μmoles n-[2-hydroxyethyl]piperazine-N'-[ethanesulfonic acid] was pre-incubated for 2 min at 30° C. Enzyme in the extraction buffer (50 μL) was then added. After the incubation continued for 15 min at 30° C., 2 mL 1 N HCl was added to stop the reaction. After 15 min at room temperature, the absorbency was measured at 355 nm. A blank containing water was used. The 5,10-methenyl-THF formed was determined using an extinction coefficient of 24,900 M-1 cm-1.

Provenance & Evidence