Highly Stable Bioluminescence-Based Fiber-Optic Sensor Using Immobilized Enzymes fromVibrio Harveyi

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Abstract
The properties of the bioluminescent enzymatic systems from V. fischeri and V.harveyi immobilized on preactivated polyamide membranes, as well as the characteristics of a fiber-optic biosensor equipped with such membranes, have been studied. Particular attention has been paid to the stability of the bioactive matrices under working conditions, since this remains a key-point for designing operational biosensors of practical use. Whatever the origin of the bioluminescent system, the microdetermination of NADH could be performed at the nanomolar level with detection limits of 2 nM and 0.3 nM with the systems from V. harveyi and V. fischeri, respectively. the use of polyethyleneglycol (PEG 600) improves the operational stability of the bi-enzymatic system from V. fischeri, but a recalibration must be carried out every ten assays. Although the immobilized system from V. harveyi exhibited a lower activity than the enzymes from V. fischeri, its excellent operational stability (100 assays performed within a day without loss in activity) makes it more suitable in designing a truly operational bioluminescence-based NADH sensor.