FIREFLY LUCIFERASE PURIFICATION USING POLYETHYLENE-GLYCOL AND DYEMATREX-ORANGE-A

Efficient measurement of adenosine triphosphate by bioluminescence depends on the quality of firefly luciferase used. A rapid purification of this enzyme is reported that permits removal of enzymes interfering in the bioluminescent reaction. The enzyme was extracted from firefly tails and precipitated with PEG 20 000, and the resulting pellet was subjected to chromatography on a Dyematrex gel (Orange A), which retains the interfering enzymes but does not bind luciferase. As shown by adenylate kinase activity determination and sodium dodecyl sulfate polyacrylamide gel electrophoretic examination of the resultant preparation, partial purification of luciferase was successful in giving a preparation without interfering enzymes.