Biocatalysis of immobilized chlorophyllase in a ternary micellar system.
The immobilization of chlorophyllase was optimized by physical adsorption on various inorganic supports, including alumina, celite, Dowex-1-chloride, glass beads and silica gel. The enzyme was also immobilized in different media, including water, Tris-HCl buffer solution and a ternary micellar system containing Tris-HCl buffer solution, hexane and surfactant. The highest immobilization efficiency (84.56%) and specific activity (0.34 mumol hydrolyzed chlorophyll mg protein-1 per min) were obtained when chlorophyllase was suspended in Tris-HCl buffer solution and adsorbed onto silica gel. The effect of different ratios of chlorophyllase to the support and the optimum incubation time for the immobilization of chlorophyllase were determined to be 1-4 and 60 min, respectively. The experimental results showed that the optimum pH and temperature for the immobilized chlorophyllase were 8.0 and 35 degrees C, respectively. The use of optimized amounts of selected membrane lipids increased the specific activity of the immobilized chlorophyllase by approximately 50%. The enzyme kinetic studies indicated that the immobilized chlorophyllase showed a higher affinity towards chlorophyll than pheophytin as substrate.
Gaffar
R
R
Department of Food Science and Agricultural Chemistry, McGill University, Québec, Canada.
Kermasha
S
S
Bisakowski
B
B
D001709:Biotechnology; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D002384:Catalysis; D002734:Chlorophyll / Q000378:metabolism; D004800:Enzymes, Immobilized; D056890:Eukaryota / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008563:Membrane Lipids; D008823:Micelles; D010674:Pheophytins / Q000378:metabolism; D013696:Temperature
D004800:Enzymes, Immobilized; D008563:Membrane Lipids; D008823:Micelles; D010674:Pheophytins; D002734:Chlorophyll; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase