title: Characterization of sol-gel entrapped chlorophyllase.
abstract: Immobilization of membrane proteins remains a challenge compared to soluble proteins. The membrane protein-chlorophyllase was successful entrapped in tetramethoxysilane (TMOS)-based sol-gel in the presence of lipid. Activity was examined against mixing rate, incubation temperature, time, substrate, acetone, and canola oil concentration. The external mass transfer of chlorophyll is not the rate-limiting step at higher mixing rates. Stability against temperature and acetone as denaturant was enhanced. In spite of the fact that an initial reaction lag phase was observed, 20% more chlorophyll was hydrolyzed, compared to reaction with free enzyme by the end of a 12 h assay. The initial lower activity demonstrated by entrapped chlorophyllase is likely due to the diffusion resistance of chlorophyll into and within the entrapment matrix. This hypothesis was substantiated by a low diffusion coefficient on the order of 10(-14) m(2)/s obtained for chlorophyll in nanoporous sol-gel particles. Pore size distribution of nanoporous wet TMOS-based sol-gel with or without protein was determined by thermoporometry. The change in pore morphology upon doping with chlorophyllase suggests that protein acts as a template during the sol-gel process.
authors.affiliation: Department of Chemical Engineering, Queen's University, Kingston, Ont., Canada.
authors.forename: Yunyu
authors.initials: Y
authors.lastname: Yi
authors.forename: Selim
authors.initials: S
authors.lastname: Kermasha
authors.forename: Ronald
authors.initials: R
authors.lastname: Neufeld
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000302:isolation & purification*; D004058:Diffusion; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005524:Food Technology; D005782:Gels; D007700:Kinetics; D012821:Silanes / Q000737:chemistry* / Q000494:pharmacology; D012995:Solubility
chemical_list: D004800:Enzymes, Immobilized; D005782:Gels; D012821:Silanes; C061205:tetramethoxysilane; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase