A Rapid Determination of ATP in Fish Muscle Using Luciferase of American Firefly Photinus pyrarlis

Abstract

The quantitative method of ATP in fish muscle was investigated by the measurement of emitted light intensity using luciferase of the American firefly. The peak of the emitted light spectrum was shown at 554nm. The sensitive assay for ATP was improved by increasing the concentration of the enzyme with the presence of bovine serum albumin, DTT, and EDTA. The regression line was obtained between the emitted light intensity and the ATP concentra-tion in the ranges from 1 to 50μM. The bioluminescent reaction was not inhibited by the dilu-tion of neutralized perchlorate as extractive solvent with 100 volumes of distilled water.
The ATP contents in the extracts of fish muscle were determined both by bioluminescent reaction and by column chromatography. The values obtained by the two different methods almost agreed. Consequently, the bioluminescent reaction of American firefly luciferase could be used for the rapid determination of the ATP contents in fish muscle.