| abstract |
The invention relates to complexes that include lipases covalently bonded, by means of regions not involved in the active centre thereof, to biomolecules of interest (proteins such as enzymes, antibodies or proteins A or G, cofactors or DNA probes). The complexes can be immobilised via the intact active centre of the lipases on all types of hydrophobic media, without requiring any modifications or activations thereof. The bonding of the lipase to the hydrophobic media is reversible, allowing the reversible immobilisation of the complexes. The invention provides two methods, chemical and biological, for preparing said complexes, as well as a method for immobilising said complexes. Said immobilised complexes can be used in industry, in laboratories or in diagnostics, given the potential use thereof as biocatalysts or as biosensors. The chemical method includes adsorbing the lipase to a porous hydrophobic medium, combining the lipase thus adsorbed with the biomolecule, and desorbing said complex from the medium using detergents. The biological method includes inserting the gene of the biomolecule in a plasmid containing the gene of the lipase, in a microorganism, in order for the latter to express the lipase fused with the biomolecule. The reversible immobilisation of the complexes generated using either of the two methods can be carried out on non-porous hydrophobic media. |