Characterization and purification of biliverdin reductase from the liver of eel, Anguilla japonica

Abstract

1. 1. Biliverdin reductase from the liver of eel, Anguilla japonica was characterized and purified with a novel enzymatic staining method on polyacrylamide electrophoretic gel. 2. 2. This enzyme could use both NADPH and NADH as coenzyme. The K m of NADPH was 5.2 μM, while that of NADH was 5.50 μM. 3. 3. The optimum reaction pH for using HADPH as coenzyme was 5.3. That for NADH was 6.1. The optimum reaction temperature is 37°C. 4. 4. When NADPH was used as coenzyme, the K m of biliverdin was 0.6 μM. When NADH was used as coenzyme, the K m of biliverdin was 7.0 μM. 5. 5. The activity of the enzyme was inhibited by the concentration of biliverdin. Also, the potency of the enzyme was much less than that of the analogous enzyme isolated from mammals. 6. 6. This is a fairly stable enzyme with a mol. wt around 67,000. Its estimated pI was pH 3.5–4.0. 7. 7. This is the first time biliverdin reductase has been isolated and characterized from a vertebrate other than mammals. The property of it is quite different from that of mammals.