title: Covalent binding of urease on ammonium-selective potentiometric membranes.
abstract: As part of the development of disposable urea bioselective probes, the covalent binding of urease on ammonium-selective potentiometric membranes has been assessed. Nonactin/bis(1-butylpentyl)adipate/poly(vinylchloride) (PVC) membranes, directly applied to an internal solid contact (conductive epoxy-graphite composite), has been used as a support for covalent immobilization of urease. Two types of all-solid-state construction process have been assayed: thin layers of cellulose acetate (CA) were coated on the PVC ammonium-selective membranes (type 1) and blends of PVC and CA at various ratios were used as ammonium-selective membrane matrices (type 2). Urease was covalently attached to CA via aldehyde groups. These groups were created on the polysaccharide with sodium periodate to which the enzyme was immobilized through a spacer (hexamethylenediamine). The viability of both types of probe for the determination of ammonium ions was assessed after each step of the activation process. Results indicated that type 2 potentiometric probes are altered after the treatment with sodium periodate. Good results were obtained with type 1 probes. Their dynamic concentration range of response to urea was from 2 x 10(-5) to 0.01 M with a sensibility of 50 mV/decade.
authors.affiliation: Department of Chemistry, University of Coimbra, Portugal.
authors.forename: M H
authors.initials: MH
authors.lastname: Gil
authors.forename: A P
authors.initials: AP
authors.lastname: Piedade
authors.forename: S
authors.initials: S
authors.lastname: Alegret
authors.forename: J
authors.initials: J
authors.lastname: Alonso
authors.forename: E
authors.initials: E
authors.lastname: Martinez-Fàbregas
authors.forename: A
authors.initials: A
authors.lastname: Orellana
mesh_terms: D000641:Ammonia; D015374:Biosensing Techniques; D002482:Cellulose / Q000031:analogs & derivatives; D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D015394:Molecular Structure; D011143:Polyvinyl Chloride; D011199:Potentiometry; D012680:Sensitivity and Specificity; D014508:Urea / Q000378:metabolism*; D014510:Urease / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; C005062:acetylcellulose; D000641:Ammonia; D014508:Urea; D011143:Polyvinyl Chloride; D002482:Cellulose; D014510:Urease