Compartmentalized system with membrane-bound glycerol kinase. Activity and product distribution versus asymmetrical substrate supply.
An artificial-membrane-bound glycerokinase chosen as a membrane-bound two-substrate-enzyme model has been used to separate two unequal compartments of a specially designed diffusion cell. An interesting feature is the asymmetry of compartments and the existence of a diffusion layer adjacent to only one face of the enzymic membrane. In such a situation the apparent enzyme activity and the product distribution in the system have been studied versus all the possibilities of combination of ATP and glycerol supply. Our approach has lead us to differentiate two different roles played by a diffusion layer adjacent to a permeable enzymic membrane. Depending on the spatial origin of the enzymic substrates (i.e. from which compartment they derive), the diffusion layer can play either the role of a passive additional resistance to that of the membrane or the role of a third compartment in which the reaction product can partially accumulate before splitting on both parts of the membrane. Our results mainly demonstrate that a membrane-bound enzyme activity and the resulting product distribution occurring in a compartmentalized system may be regulated by the cumulative effect due to the asymmetry in volumes of the compartments, the presence of a diffusion layer and the different possibilities of substrate supply. With the topography studied, which is close to that reported for many 'in vivo' situations, the product may be diffused lead to vectorial metabolism processes.
Girard
A
A
Laboratoire de Génie Enzymatique, Atelier de Biotechnologie, (CNRS-Université Claude Bernard Lyon 1), Villeurbanne, France.
Merchie
B
B
Maïsterrena
B
B
D004800:Enzymes, Immobilized; D004926:Escherichia coli / Q000201:enzymology; D005991:Glycerol Kinase / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D013379:Substrate Specificity
D004800:Enzymes, Immobilized; D005991:Glycerol Kinase