Immobilization of tyrosinase on poly(indole-5-carboxylic acid) evidenced by electrochemical and spectroscopic methods.
A conducting, polymeric film of poly(indole-5 carboxylic acid) has been prepared by electrochemical polymerization for covalent immobilization of an enzyme belonging to the family of phenoloxidases-tyrosinase. The polymer was characterized by cyclic voltammetry, UV-VIS and Raman spectroscopy in a buffer solution. As the polymer contains pendant carboxylic groups one-step carbodiimide method was used to immobilize tyrosinase on the polymer matrix. Immobilization of tyrosinase was confirmed by surface enhanced resonance Raman scattering spectra (SERRS) and by cyclic voltammetry as well. Tyrosinase was shown to retain its biological activity when being immobilized on the polymer surface. As proved by the electrochemical and spectroelectrochemical (UV-VIS) experiments, tyrosinase covalently bonded to the polymer matrix effectively catalyzes oxidation of catechol. The reduction current of o-quinones was measured as a function of catechol concentration. The linear dependence was found to be 15 microM of catechol with sensitivity of 250 mA/M cm2.
Biegunski
A T
AT
Faculty of Chemistry, Warsaw University 02-093 Warsaw, Pasteur 1, Poland.
Michota
A
A
Bukowska
J
J
Jackowska
K
K
D002384:Catalysis; D002396:Catechols / Q000737:chemistry; D004563:Electrochemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D007211:Indoles / Q000138:chemical synthesis / Q000737:chemistry*; D014442:Monophenol Monooxygenase / Q000737:chemistry*; D010084:Oxidation-Reduction; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D011809:Quinones / Q000138:chemical synthesis / Q000737:chemistry; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013499:Surface Properties
D002396:Catechols; D004800:Enzymes, Immobilized; D007211:Indoles; D011108:Polymers; D011809:Quinones; C514680:poly(indole-5-carboxylic acid); D014442:Monophenol Monooxygenase; C034221:catechol