title: Use of reversible denaturation for adsorptive immobilization of urease.
abstract: Urease was chosen as a model multimeric protein to investigate the utility of reversible denaturation for immobilization to a hydrophobic support. Of the various procedures investigated, acidic denaturation provided the highest degree of immobilization and enzymatic activity with lowering of Km (apparent). Exposure of hydrophobic clusters in the protein molecule induced by the acidic pH environment was confirmed by fluorescence studies using 8-anilino-1-naphtalene-sulfonate as a hydrophobic-reporter probe. The catalytic potential of the enzyme at low pH values was dramatically improved with significant heat and pH stability enhancement on immobilization. Furthermore, the immobilized preparation was used successfully in continuous catalytic transformations. Based on the results presented in this article and a recent report involving a relatively more simple monomeric protein, it is suggested that reversible denaturation may be of general utility for immobilization of proteins, which are not normally adsorbed on hydrophobic supports.
authors.affiliation: Institute of Biochemistry and Biophysics, University of Tehran, Iran.
authors.forename: F
authors.initials: F
authors.lastname: Azari
authors.forename: S
authors.initials: S
authors.lastname: Hosseinkhani
authors.forename: M
authors.initials: M
authors.lastname: Nemat-Gorgani
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D002384:Catalysis; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007887:Fabaceae / Q000201:enzymology; D005456:Fluorescent Dyes / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D011489:Protein Denaturation / Q000502:physiology; D012685:Sepharose / Q000031:analogs & derivatives*; D013696:Temperature; D014510:Urease / Q000378:metabolism*
chemical_list: D000817:Anilino Naphthalenesulfonates; D004800:Enzymes, Immobilized; D005456:Fluorescent Dyes; C034543:palmityl-sepharose; C027132:1-anilino-8-naphthalenesulfonate; D012685:Sepharose; D014510:Urease