Encapsulation of enzymes in liposomes: high encapsulation efficiency and control of substrate permeability.
Enzyme encapsulation into liposomes is a promising technique to stabilize and prevent them from denaturation and proteolysis. We demonstrate this using acetylcholinesterase which is the main target for pesticides. In order to achieve a reasonable encapsulation yield, we analyzed the parameters involved in each step of various encapsulation procedures. The only encapsulation method which did not denature the protein was the lipid film hydration technique, however the encapsulation efficiency was usually low. The efficiency could be increased up to more than 40% by induction of a specific interaction between the enzyme and the lipid surface. Once encapsulated, the enzyme encountered another problem: the permeability barrier of the lipid membrane drastically diminished the activity of the enzyme entrapped in the liposome by reducing the entrance rate of the substrate molecules and then reducing the substrate concentration inside the liposome. To solve this problem, we controlled the permeability of the liposome wall by reconstituting a porin from Escherichia coli. We succeeded to recover the full functionality of the enzyme, while retaining the protection against denaturation and proteolytic enzymes.
Chaize
Barnabé
B
Groupe de Biophysique et de Biotechnologie des Protéines, IPBS-UMR 5089, Toulouse, France.
Colletier
Jacques-Philippe
JP
Winterhalter
Mathias
M
Fournier
Didier
D
D000110:Acetylcholinesterase / Q000008:administration & dosage / Q000378:metabolism; D000818:Animals; D029721:Drosophila Proteins; D004337:Drug Carriers / Q000737:chemistry*; D004795:Enzyme Stability; D004798:Enzymes / Q000008:administration & dosage* / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D010539:Permeability; D018272:Porins / Q000494:pharmacology
D029721:Drosophila Proteins; D004337:Drug Carriers; D004798:Enzymes; D008081:Liposomes; D018272:Porins; D000110:Acetylcholinesterase