Catalase-conjugated liposomes encapsulating glucose oxidase for controlled oxidation of glucose with decomposition of hydrogen peroxide produced.
The catalase-conjugated liposome encapsulating glucose oxidase (CLG) was prepared for developing a novel liposomal system for glucose oxidation with controllable enzyme activities. The catalase molecules were conjugated to the surface of liposome with 100 nm in mean diameter through coupling with the membrane-incorporated 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(glutaryl) (NGPE) at its mole fraction f(G) of 0.05 or 0.15. The average number of enzyme molecules per CLG with f(G) of 0.15 was 8.7 for glucose oxidase and 6.5 for catalase. The CLG-catalyzed oxidation of glucose was performed at 40 degrees C for prolonged period up to 99 h. The CLG with f(G) of 0.15 gave larger oxidation rate than that with f(G) of 0.05. In the fed-batch oxidation of glucose catalyzed by the former CLG, the stable oxidation rate was observed for 75 h with negligible accumulation of H(2)O(2) produced because of the durable catalytic actions of the liposomal enzymes. The oxidation rate of the CLG reaction increased to 1.1 mM-glucose/(hmM-lipid) at the acidic pH in the internal phase of liposome and the neutral pH in the external one corresponding to the optimal pH conditions for the activities of glucose oxidase and catalase, respectively. The oxidation rate catalyzed by the CLG could be controlled by adding sublytic concentrations of cholate to increase permeability of the liposome membrane to glucose. The catalase-conjugated liposomal system is potentially utilized for controlling the rate of reactions catalyzed by a variety of oxidases.
Department of Applied Molecular Bioscience, Yamaguchi University, 2-16-1 Tokiwadai, Ube 755-8611, Japan. yosimoto@yamaguchi-u.ac.jp
Makoto
M
Yoshimoto
Noriyuki
N
Takaki
Miku
M
Yamasaki
D000818:Animals; D001234:Aspergillus niger / Q000201:enzymology*; D055162:Biocatalysis / Q000187:drug effects; D002374:Catalase / Q000378:metabolism*; D002417:Cattle; D020355:Cholates / Q000494:pharmacology; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D005949:Glucose Oxidase / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D009391:Nephelometry and Turbidimetry; D010084:Oxidation-Reduction / Q000187:drug effects; D013997:Time Factors
D020355:Cholates; D004800:Enzymes, Immobilized; D008051:Lipid Bilayers; D008081:Liposomes; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase; D002374:Catalase; D005947:Glucose