Glutaraldehyde activated eggshell membrane for immobilization of tyrosinase from Amorphophallus companulatus: application in construction of electrochemical biosensor for dopamine.

Tyrosinase from a plant source Amorphophallus companulatus was immobilized on eggshell membrane using glutaraldehyde. Among the three different approaches used for immobilization, activation of eggshell membrane by glutaraldehyde followed by enzyme adsorption on activated support could stabilize the enzyme tyrosinase and was found to be effective. K(m) and V(max) values for dopamine hydrochloride calculated from Lineweaver-Burk plot were 0.67 mM and 0.08 mM min(-1), respectively. Studies on effect of pH showed retention of more than 90% activity over a pH range 5.0-6.5. Membrane bound enzyme exhibited consistent activity in the temperature range 20-45 degrees C. Shelf life of immobilized tyrosinase system was found to be more than 6 months when stored in phosphate buffer at 4 degrees C. An electrochemical biosensor for dopamine was developed by mounting the tyrosinase immobilized eggshell membrane on the surface of glassy carbon electrode. Dopamine concentrations were determined by the direct reduction of biocatalytically liberated quinone species at -0.19 V versus Ag/AgCl (3M KCl). Linearity was observed within the range of 50-250 microM with a detection limit of 25 microM.

[1]  J P Hervás Pérez,et al.  Amperometric tyrosinase biosensor based on polyacrylamide microgels. , 2006, Biosensors & bioelectronics.

[2]  Guo-Li Shen,et al.  A mediator-free phenol biosensor based on immobilizing tyrosinase to ZnO nanoparticles. , 2006, Analytical biochemistry.

[3]  Xiaoya Hu,et al.  Determination of cyanide using a tyrosinase amperometric biosensor with catechol as substrate , 1995 .

[4]  V. Balcão,et al.  Hydrolysis of whey proteins by proteases extracted from Cynara cardunculus and immobilized onto highly activated supports. , 2001, Enzyme and microbial technology.

[5]  S. Tükel,et al.  Immobilization and kinetics of catalase onto magnesium silicate , 2004 .

[6]  M. Karve,et al.  Electrochemical biosensor for catechol using agarose-guar gum entrapped tyrosinase. , 2007, Journal of biotechnology.

[7]  Horace Traubel.  Collect , 1904 .

[8]  Silvia Fabiano,et al.  Amperometric tyrosinase based biosensor using an electrogenerated polythiophene film as an entrapment support. , 2003, Talanta.

[9]  M. Furusawa,et al.  Flow-injection determination of l-tyrosine in serum with immobilized tyrosinase. , 1993, Talanta: The International Journal of Pure and Applied Analytical Chemistry.

[10]  Keiichi Kaneto,et al.  Amperometric tyrosinase based biosensor using an electropolymerized PTS-doped polypyrrole film as an entrapment support , 2004 .

[11]  Martin M. F. Choi,et al.  A biosensor with myrosinase and glucose oxidase bienzyme system for determination of glucosinolates in seeds of commonly consumed vegetables , 2005 .

[12]  Martin M. F. Choi,et al.  A homocysteine biosensor with eggshell membrane as an enzyme immobilization platform , 2006 .

[13]  S. F. D’souza,et al.  Immobilization and stabilization of biomaterials for biosensor applications , 2001, Applied biochemistry and biotechnology.

[14]  M. Karve,et al.  Characterization of tyrosinase and accompanying laccase from Amorphophallus campanulatus. , 2003, Indian Journal of Biochemistry & Biophysics.

[15]  M. Karve,et al.  Development of electrochemical biosensor based on tyrosinase immobilized in composite biopolymeric film. , 2006, Analytical biochemistry.

[16]  S. F. D’souza,et al.  IMMOBILIZED ENZYMES IN BIOPROCESS , 1999 .

[17]  Y. Yagcı,et al.  Immobilization of tyrosinase in polysiloxane/polypyrrole copolymer matrices. , 2005, International journal of biological macromolecules.

[18]  Dipali S Bagal,et al.  Entrapment of plant invertase within novel composite of agarose-guar gum biopolymer membrane , 2006 .

[19]  M. L. Mena,et al.  Development of a tyrosinase biosensor based on gold nanoparticles-modified glassy carbon electrodes: Application to the measurement of a bioelectrochemical polyphenols index in wines , 2005 .

[20]  Martin M. F. Choi,et al.  Immobilization of beef liver catalase on eggshell membrane for fabrication of hydrogen peroxide biosensor , 2004 .

[21]  D. Leech,et al.  Reagentless Tyrosinase Enzyme Electrodes: Effects of Enzyme Loading, Electrolyte pH, Ionic Strength, and Temperature , 1997 .

[22]  Guo-Li Shen,et al.  A phenol biosensor based on immobilizing tyrosinase to modified core-shell magnetic nanoparticles supported at a carbon paste electrode , 2005 .

[23]  C. S. Caruso,et al.  Determination of epinephrine and dopamine in pharmaceutical formulations using a biosensor based on carbon paste modified with crude extract of cara root (Dioscorea bulbifera) , 1999 .

[24]  Itaru Honma,et al.  Electrochemical biosensor based on protein-polysaccharide hybrid for selective detection of nanomolar dopamine metabolite of 3,4-dihydroxyphenylacetic acid (DOPAC) , 2005 .

[25]  Joseph Wang,et al.  Hydrazine Detection Using a Tyrosinase-Based Inhibition Biosensor , 1995 .

[26]  O. Fatibello‐Filho,et al.  A zucchini-peroxidase biosensor applied to dopamine determination. , 2005, Farmaco.

[27]  R. S. Kelly,et al.  Mediated electrochemical reduction of cytochrome c and tyrosinase at perfluorosulfonated ionomer coated electrodes , 1993 .

[28]  Xiao Dong Chen,et al.  Immobilization of β-galactosidase on graphite surface by glutaraldehyde , 2001 .

[29]  L. Macholán,et al.  Enzyme electrode with immobilized polyphenol oxidase for determination of phenolic substrates , 1977 .

[30]  Alexander M. Klibanov,et al.  Regioselective oxidation of phenols catalyzed by polyphenol oxidase in chloroform , 1985 .

[31]  D. Paolucci-Jeanjean,et al.  Immobilization of lipase on a ceramic membrane: activity and stability , 2004 .

[32]  A. Turner,et al.  Gas-Phase Microbiosensor for Monitoring Phenol Vapor at ppb Levels , 1995 .

[33]  Dan Xiao,et al.  An optical glucose biosensor with eggshell membrane as an enzyme immobilisation platform , 2001 .

[34]  Jenny Emnéus,et al.  Improved stability and altered selectivity of tyrosinase based graphite electrodes for detection of phenolic compounds , 1999 .

[35]  Martin M. F. Choi,et al.  Development and analytical application of an uric acid biosensor using an uricase-immobilized eggshell membrane. , 2007, Biosensors & bioelectronics.

[36]  Won-Yong Lee,et al.  Amperometric phenol biosensor based on sol–gel silicate/Nafion composite film , 2003 .

[37]  J. Kong,et al.  A sensitive mediator-free tyrosinase biosensor based on an inorganic-organic hybrid titania sol-gel matrix , 2003 .

[38]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.