Catalytic properties of variously immobilized mushroom tyrosinase: A kinetic study for future development of biomimetic amperometric biosensors

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388955%3A_____%2F20%3A00524213" target="_blank" >RIV/61388955:_____/20:00524213 - isvavai.cz</a>

Alternative codes found

RIV/00216275:25310/20:39916069 RIV/00216208:11110/20:10411636

Result on the web

<a href="http://hdl.handle.net/11104/0308593" target="_blank" >http://hdl.handle.net/11104/0308593</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.jelechem.2020.114066" target="_blank" >10.1016/j.jelechem.2020.114066</a>

Result language

angličtina

Original language name

Catalytic properties of variously immobilized mushroom tyrosinase: A kinetic study for future development of biomimetic amperometric biosensors

Original language description

Mushroom tyrosinase was immobilized by direct embedding into electrode material (modified carbon paste electrode), incorporation of cross-linked enzyme aggregates into a polymer membrane (glassy carbon electrode covered by thin layer of Nafion (R)), and covalent attachment using self-assembled monolayers (gold electrode with the chemically bound enzyme). Both, standard UV-Vis spectrophotometry and amperometry in a batch configuration are presented as complementary methods to study the tyrosinase enzyme kinetics, whose catecholase activity results in electroactive products (ortho-quinones). Due to higher sensitivity of amperometric detection, evident advantage in the enzyme consumption was obtained. Prepared amperometric tyrosinase biosensors were characterized using cyclic voltammetry and atomic force microscopy. The Michaelis constant values of immobilized and unbound tyrosinase (free enzyme solution) towards dopamine and catechol were compared. The apparent Michaelis constant values for immobilized tyrosinase are significantly lower than the declared value of 0.840 mmol L-1 dopamine for the unbound enzyme. The enzymetyrosinase arranged in self-assembledmonolayer serves as an efficient sensor due to lowapparent Michaelis constant of 0.061 mmol L-1 dopamine and high maximum reaction velocity of 0.458 mu A s(-1). This fact reflects the ideal arrangement of enzymemolecules causing high availability of the binding site. Tris-glycine sodiumdodecyl sulphate polyacrylamide gel electrophoresis and atomic force microscopy clarified that the protein of molecular weight 25 kDa is bound preferably on chemically modified gold electrode. A sensor prepared by the immobilization of tyrosinase on gold electrode results in higher catecholase activity towards dopamine than in case of CPE and GC electrodes, where enzyme is immobilized physically.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10403 - Physical chemistry

Project

<a href="/en/project/GA19-03160S" target="_blank" >GA19-03160S: Electrochemical study on new artificial enzymes and their role in sensing of neurotransmitters</a><br>

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2020

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Electroanalytical Chemistry

ISSN

1572-6657

e-ISSN

—

Volume of the periodical

864

Issue of the periodical within the volume

MAY 2020

Country of publishing house

CH - SWITZERLAND

Number of pages

9

Pages from-to

114066

UT code for WoS article

000528254900003

EID of the result in the Scopus database

2-s2.0-85082804107