Journal ArticleDOI
Collagen strip with immobilized luciferase for ATP bioluminescent determination.
TLDR
FireflyLuciferase from Photinus pyralis has been covalently bound to a collagen strip via an acylazide activation process and the results were found in good agreement with those obtained by soluble luciferase.Abstract:
Firefly luciferase from Photinus pyralis has been covalently bound to a collagen strip via an acylazide activation process. Immobilization performed in the presence of both substrates ATP and luciferin allows to increase the activity retained on the strip. The best activity exhibited by immobilized luciferase was obtained in a 0.05M Tris-acetate buffer, pH 7.75. The pH optimum and the activation energy of luciferase have been found unchanged after immobilization. In the chosen stirring conditions, no diffusional limitations of substrates appear. ATP measurements can be performed with collagen-bound luciferase in the range 1.10−11M−3.10−6M. It was possible to store the strips at 4°C in a dehydrated form; then, the bound enzyme retains 20% of its initial activity after eight months. Human blood ATP was measured with this collagen-bound luciferase and the results were found in good agreement with those obtained by soluble luciferase.read more
Citations
More filters
Book ChapterDOI
[2] A survey of enzyme coupling techniques
TL;DR: No single immobilization method is best for all enzymes or all applications of any given enzyme, so a survey of enzyme coupling techniques and enzyme immobilization, with emphasis on the carder matrices, is discussed.
Journal ArticleDOI
Methods for the determination of adenosine triphosphate and other adenine nucleotides
TL;DR: The following methods for the determination of adenosine triphosphate reported in the past 25 years are considered: bioluminescence methods with the use of the firefly luciferase enzyme, and fluorescence, spectrophotometric, and electrochemical techniques, which are promising but not commonly used for thedetermination of adenine nucleotides.
Journal ArticleDOI
Ultrasensitive ATP detection using firefly luciferase entrapped in sugar-modified sol-gel-derived silica.
TL;DR: Firefly luciferase was entrapped in sol-gel-derived silica containing precursors based on covalent linkage of d-gluconolactone or d-maltonolact one to form N-(3-triethoxysilylpropyl)glu Conamide or N-( 3-triETHoxysilane)maltonamide and showed catalytic constants close to those in solution.
Journal ArticleDOI
Luminescence Fiber-Optic Biosensor
TL;DR: A novel type of biosensors involving immobilized bioluminescence enzymes and a fiber-optic probe has been developed and the reaction catalyzed by immobilized horseradish peroxidase has been used for hydrogen peroxide determination.
Journal ArticleDOI
Fibre‐optic biosensor based on luminescence and immobilized enzymes: Microdetermination of sorbitol, ethanol and oxaloacetate
TL;DR: This work has investigated highly selective and ultrasensitive biosensors based on luminescent enzyme systems linked to optical transducers and used them for the microdetermination of sorbitol, ethanol and oxaloacetate at the nanomolar level with a good precision.
References
More filters
Journal ArticleDOI
Continuous monitoring of ATP-converting reactions by purified firefly luciferase
TL;DR: The continuous monitoring of ATP concentration by firefly luciferase was used for kinetic determination of enzymes and metabolites and for endpoint analysis of metabolites and was found to be extremely sensitive and convenlent for routine applications.
Journal ArticleDOI
Kinetics of the firefly luciferase catalyzed reactions
Journal ArticleDOI
Enzyme collagen membrane for electrochemical determination of glucose
TL;DR: In this paper, a new trace glucose analyzer has been designed using electrochemical sensors, which includes a glucose sensor consisting of a modified gas electrode in which pH detector was replaced by a platinum disk and the porous film by a collagen membrane on which beta-D-glucose oxidase has been covalently bound after an acyl-azide activation process.
Journal ArticleDOI
A mild method of general use for covalent coupling of enzymes to chemically activated collagen films
TL;DR: Films of highly polymerized collagen, prepared in industrial conditions, were chosen for surface covalent binding of enzymes because of their insolubility, mechanical resistance, proteic nature, hydrophilic properties and for their abundance in chemically activable COOH.
Related Papers (5)
Immobilization of firefly luciferase on glass rods: properties of the immobilized enzyme.
Bioluminescent assays with immobilized firefly luciferase based on flow injection analysis
Paul J. Worsfold,Abdul Nabi +1 more