[Immobilized luciferase of Luciola mingrelica fireflies. The kinetic properties and thermostability of luciferase immobilized on cellulose films].
Luciferase of fireflies Luciola mingrelica was immobilized on cellulose films activated by cyanuric chloride or sodium periodate. Kinetic properties and the contribution of diffusional obstacles to the kinetics of the immobilized enzyme were examined. External and internal diffusion were found to influence the kinetic parameters. The stability of the enzyme was investigated at 25 degrees C and pH 7.8. Thermoactivation of the immobilized enzyme was shown to proceed in two stages: fast and slow. Dithiotreitol and cystein stabilized the enzyme at the fast stage while salt supplements at both stages. The fast thermoinactivation stage was apparently associated with the oxidation of luciferase SH-groups. It is demonstrated that the immobilized enzyme of Luciola mingrelica can be employed to measure ATP traces with the detection limit 0.1 mM. The enzyme immobilized on cellulose films can be used repeatedly.
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002482:Cellulose / Q000494:pharmacology*; D001517:Coleoptera / Q000201:enzymology*; D004058:Diffusion; D004355:Drug Stability; D004800:Enzymes, Immobilized / Q000494:pharmacology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000494:pharmacology; D007700:Kinetics; D008156:Luciferases / Q000494:pharmacology*
D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D000255:Adenosine Triphosphate; D002482:Cellulose; D008156:Luciferases
Immobilizovannaia liutsiferaza svetliakov Luciola mingrelica. Kineticheskie svoĭstva i termostabil'nost' liutsiferazy, immobilizovannoĭ na tselliuloznykh plenkakh.