title: Immobilization of the urease on eggshell membrane and its application in biosensor.
abstract: Eggshell membrane is a natural material, essentially made up of protein fibers having flexibility in the aqueous solution and possessing gas and water permeability. It is used as a biomembrane for immobilization of urease for the development of a potentiometric urea biosensor. Eggshell membrane was treated with polyethyleneimine (PEI) to impart polycation characteristics. Urease was immobilized on the PEI treated eggshell membrane through adsorption. SEM study was carried out to observe the changes in surface morphology after immobilization. FTIR study of membrane was carried out to observe the changes in IR spectra after immobilization of enzyme. Immobilized membrane was associated with ammonium ion selective electrode. Biosensor exhibited sigmoidal responses for the urea concentration range from 0.5 to 10mM. The response time of the biosensor was 120 s. A single membrane was reused for 270 reactions without loss of activity. The urease-eggshell membranes were stable for 2 months when stored in buffer even at room temperature.
authors.affiliation: Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre Trombay, Mumbai-400 085, India. Electronic address: sfdsouza@barc.gov.in.
authors.forename: S F
authors.initials: SF
authors.lastname: D'Souza
authors.affiliation: Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre Trombay, Mumbai-400 085, India. Electronic address: jkumar@barc.gov.in.
authors.forename: Jitendra
authors.initials: J
authors.lastname: Kumar
authors.affiliation: Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre Trombay, Mumbai-400 085, India.
authors.forename: Sandeep Kumar
authors.initials: SK
authors.lastname: Jha
authors.affiliation: Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre Trombay, Mumbai-400 085, India.
authors.forename: B S
authors.initials: BS
authors.lastname: Kubal
mesh_terms: D015374:Biosensing Techniques; D002462:Cell Membrane / Q000737:chemistry; D004531:Eggs; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D017736:Ion-Selective Electrodes; D008855:Microscopy, Electron, Scanning; D011094:Polyethyleneimine / Q000737:chemistry; D011199:Potentiometry; D017550:Spectroscopy, Fourier Transform Infrared; D014508:Urea / Q000032:analysis; D014510:Urease / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D014508:Urea; D011094:Polyethyleneimine; D014510:Urease