MEDIATED AMPEROMETRIC ASSESSMENT OF ENZYME ACTIVITY IN LIVING S. CEREVISIAE CELLS IMMOBILIZED ON PLATINUM MICRO-BAND ELECTRODE CHIP

摘要

Screening systems capable of assessing enzyme activity in whole living cells, can produce information related to i) the function of enzymes in the native environment and ii) the cellular cofactors [1] as well as iii) the cellular signaling and regulation mechanisms, together forming the functional basis of the organization of the cellular reaction pathways, i.e. metabolons [2]. Since it has been indicated that the cellular redox environment is related to e.g. the pathogenesis of neurodegenerative diseases [3], it is of utmost importance to develop methods capable of assessing intracellularly the activity of redox enzymes. Considering the low concentrations of the compounds to be detected, it is necessary to miniaturize the electrochemical system making the cell-electrode distance as small as possible. In order to realize this goal, a mediated amperometric method, employing microelectrodes in chip-format, was developed for assessing the activity of intracellular redox enzymes.