Purification and characterization of luciferases from fireflies, Luciola cruciata and Luciola lateralis.
Luciferases of Luciola cruciata and Luciola lateralis, LcL and LlL, were purified to homogeneity by ammonium sulfate precipitation, gel-filtration column chromatography, and hydroxyapatite HPLC. The molecular masses of the enzymes determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were both 62 kDa, almost identical to that of Photinus pyralis (PpL). LcL was found to be similar to PpL in thermal stability, pH stability, and the wavelength of maximum light intensity. LlL was superior to LcL and PpL in thermal and pH stability, and the reaction catalyzed by LlL emits green light with a peak intensity at 552 nm, which is 10 nm shorter in wavelength than those of PpL and LcL.
Kajiyama
N
N
Research and Development Division, Kikkoman Corporation, Chiba, Japan.
Masuda
T
T
Tatsumi
H
H
Nakano
E
E
D000818:Animals; D001517:Coleoptera / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008156:Luciferases / Q000737:chemistry / Q000302:isolation & purification*; D008163:Luminescent Measurements; D008970:Molecular Weight; D013057:Spectrum Analysis
D008156:Luciferases