12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Evaluation of the affinity of bacterial luciferases for immobilization on hydrophobic supports

Hosseinkhani, Saman^1,2, Szittner, Rose¹, Nemat-Gorgani, Mohsen², Meighen, Edward¹

1. Dept of Biochemistry, McGill University, Montreal, PQ, Canada

Email: meighen@med.mcgill.ca

2. Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran

Immobilized enzymes are widely used for clinical and industrial purposes for the assay of different analytes and enzymes. The use of immobilization is very advantageous as the bound enzyme can easily be recovered and reused and in many cases its stability is greater than that of the soluble enzyme. Bacterial luciferases can be readily applied to measure a variety of different compounds particularly those whose oxidation can be coupled with the production of NAD(P)H and/or FMNH₂. A simple method involving fluorescent markers was applied to screen different bacterial luciferases for their affinity to hydrophobic matrices before measuring the binding of luciferase to the matrix. As a high stability and activity for luciferase is very desirable for analytical assays, the kinetic and physical properties of luciferases immobilized on hydrophobic supports containing alkyl chains of varying length were determined. These results indicate that this approach can lead to active and stable immobilized luciferases that can rapidly be prepared and reutilized in light-emitting assays (Supported by grant, MT-4314 from the Canadian Institutes of Health Research [CIHR]).

This is a preprint of an article accepted for publication in Luminescence: Copyright 2001 John
Wiley & Sons, Ltd (Wiley website)