Improvement of thermostability and activity of firefly luciferase through [TMG][Ac] ionic liquid mediator.
Firefly luciferase catalyzes production of light from luciferin in the presence of Mg(2+)-ATP and oxygen. This enzyme has wide range of applications in biotechnology and development of biosensors. The low thermal stability of wild-type firefly luciferase is a limiting factor in most applications. Improvements in activity and stability of few enzymes in the presence of ionic liquids were shown in many reports. In this study, kinetic and thermal stability of firefly luciferase from Photinus pyralis in the presence of three tetramethylguanidine-based ionic liquids was investigated. The enzyme has shown improved activity in the presence of [1, 1, 3, 3-tetramethylguanidine][acetate], but in the presence of [TMG][trichloroacetate] and [TMG][triflouroacetate] activity, it decreased or unchanged significantly. Among these ionic liquids, only [TMG][Ac] has increased the thermal stability of luciferase. Incubation of [TMG][Ac] with firefly luciferase brought about with decrease of K(m) for ATP.
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. mehd_ebrahimi@yahoo.com
Mehdi
M
Ebrahimi
Saman
S
Hosseinkhani
Akbar
A
Heydari
Ramazan Ali
RA
Khavari-Nejad
Jafar
J
Akbari
D000818:Animals; D004795:Enzyme Stability; D048888:Fireflies / Q000737:chemistry / Q000201:enzymology*; D006146:Guanidines / Q000737:chemistry*; D006358:Hot Temperature; D019476:Insect Proteins / Q000737:chemistry* / Q000378:metabolism; D052578:Ionic Liquids / Q000737:chemistry*; D007700:Kinetics; D049409:Luciferases, Firefly / Q000737:chemistry* / Q000378:metabolism
D006146:Guanidines; D019476:Insect Proteins; D052578:Ionic Liquids; D049409:Luciferases, Firefly; C477069:1,1,3,3-tetramethylguanidine