title: Kinetic and circular dichroism studies of enzymes adsorbed on ultrafine silica particles.
abstract: Negatively charged ultrafine silica particles (average diameter 20 nm) were used as support materials for adsorption immobilization of porcine trypsin, horseradish peroxidase, and bovine catalase under various conditions, and the changes in the enzyme activities and the circular dichroism (CD) spectra of these enzymes upon adsorption were measured. Since the light scattering intensity of the ultrafine particles was very low, the activities and the CD spectra of the enzymes adsorbed on the particle surfaces could be measured. The enzymes adsorbed at pH around and above their isoelectric points (pI) showed high activities. On the other hand, the enzymes adsorbed at pHs below their pI had significantly diminished activities and showed large CD spectral changes upon adsorption. The extent of CD spectral changes in the enzymes upon adsorption correlated very closely with that of the activity reduction. Therefore, the conformational changes in enzymes upon adsorption are one of the important factors that reduce the activities of adsorbed enzymes. These results demonstrate that the ultrafine particles are not only a novel support for enzyme immobilization but also are helpful for the molecular understanding of the immobilized enzymes.
authors.affiliation: Applied Chemistry Department, Kyushu Institute of Technology, Kitakyushu, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Kondo
authors.forename: F
authors.initials: F
authors.lastname: Murakami
authors.forename: M
authors.initials: M
authors.lastname: Kawagoe
authors.forename: K
authors.initials: K
authors.lastname: Higashitani
mesh_terms: D000327:Adsorption; D000818:Animals; D002374:Catalase / Q000378:metabolism; D002417:Cattle; D002942:Circular Dichroism; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D012822:Silicon Dioxide; D013552:Swine; D014357:Trypsin / Q000378:metabolism
chemical_list: D004800:Enzymes, Immobilized; D012822:Silicon Dioxide; D006735:Horseradish Peroxidase; D002374:Catalase; D014357:Trypsin