A new ultrasensitive bioluminogenic enzyme substrate for beta-galactosidase.
A derivative of D-luciferin, D-luciferin-O-beta-galactoside, was synthesized and used as highly sensitive substrate for beta-galactosidase. The substrate was physicochemically characterized. Enzymatic cleavage of the new compound by beta-galactosidase was demonstrated and kinetic constants Km, Vmax, kcat and kcat/Km have been determined. The compound has been proved to be a highly sensitive substrate for beta-galactosidase, permitting a limit of detection of 3.7 x 10(-19) mol of enzyme per assay.
Geiger
R
R
Medor GmbH, Herrsching, Germany.
Schneider
E
E
Wallenfels
K
K
Miska
W
W
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000201:enzymology; D049454:Firefly Luciferin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D005697:Galactosides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007536:Isomerism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D001616:beta-Galactosidase / Q000378:metabolism*
D005697:Galactosides; C080091:luciferin-O-galactopyranoside; D049454:Firefly Luciferin; D001616:beta-Galactosidase