Immobilization and biocatalysis of chlorophyllase in selected organic solvent systems.
Chlorophyllase extract from Phaeodactylum tricornutum was immobilized by physical adsorption on DEAE-cellulose and silica gel as well as by covalent binding on Eupergit C, Eupergit C250L, Eupergit C/ethylenediamine (EDA) and Eupergit C250L/EDA. Although the highest immobilization yield (83-93%) and efficiency (51-53%) were obtained when chlorophyllase extract was immobilized on DEAE-cellulose and silica gel, there was no improvement in the thermal stability of chlorophyllase as compared to that of the free one. The immobilization of chlorophyllase extract on Eupergit C250L/EDA resulted by a high recovery of enzymatic activity, with an immobilization efficiency of 44%, and promoted a higher stabilization of chlorophyllase (four times) in the aqueous/miscible organic solvent medium. On the other hand, the inhibitory effect of refined bleached deodorized (RBD) canola oil was reduced by immobilization of chlorophyllase extract onto silica gel as compared to those obtained with other enzyme preparations. However, the re-cycled chlorophyllase extract immobilized on Eupergit C250L/EDA retained more than 75% of its initial enzyme activity after 6 cycles, whereas that immobilized on silica gel was completely inactivated. The highest catalytic efficiency, for both free and immobilized chlorophyllase on Eupergit C250L/EDA, was obtained in the ternary micellar system as compared to the aqueous/miscible organic solvent and biphasic media.
Karboune
Salwa
S
Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore, Ste-Anne de Bellevue, Quebec, Canada H9X 3V9.
Neufeld
Ronald
R
Kermasha
Selim
S
D000327:Adsorption; D001709:Biotechnology; D002265:Carboxylic Ester Hydrolases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D002384:Catalysis; D002734:Chlorophyll / Q000032:analysis / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005029:Ethylenediamines / Q000737:chemistry*; D056890:Eukaryota / Q000201:enzymology; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry / Q000494:pharmacology; D007700:Kinetics; D008823:Micelles; D010936:Plant Extracts / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding; D000074262:Rapeseed Oil; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry* / Q000145:classification; D013696:Temperature; D014867:Water / Q000737:chemistry
D004800:Enzymes, Immobilized; D005029:Ethylenediamines; D005229:Fatty Acids, Monounsaturated; D008823:Micelles; D010936:Plant Extracts; D011108:Polymers; D000074262:Rapeseed Oil; D012997:Solvents; D014867:Water; D002734:Chlorophyll; D058428:Silica Gel; D012822:Silicon Dioxide; C065843:Eupergit C; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase