Khlupova Maria, Kuznetsov Boris, Demkiv Olha, Gonchar Mykhailo, Csöregi Elisabeth, Shleev Sergey
Laboratory of Chemical Enzymology, A.N. Bach Institute of Biochemistry, 119071 Moscow, Russia.
Talanta. 2007 Feb 15;71(2):934-40. doi: 10.1016/j.talanta.2006.05.069. Epub 2006 Jul 10.
Intact and permeabilized yeast cells were tested as the biorecognition elements for amperometric assay of formaldehyde (FA). For this aim, the mutant C-105 (gcr1 catX) of the methylotrophic yeast Hansenula polymorpha with a high activity of AOX was chosen. Different approaches were used for monitoring FA-dependent cell response including analysis of their oxygen consumption rate by the use of a Clark electrode, as well as assay of oxidation of redox mediator at a screen-printed platinum electrode covered by cells entrapped in Ca-alginate gel. It was shown that oxygen consumption rate of permeabilized cells reached its saturation at 4mM of FA (23 degrees C). The detection limit was found to be 0.27mM. In the presence of redox mediator 2,6-dichlorophenolindophenol (DCIP), the screen-printed platinum band electrode covered by permeabilized cells did not show any current output to FA. In contrast, well-pronounced amperometric response to FA was observed in the case of intact yeast cells in the presence of DCIP. It was shown that current output reached its maximum at 7mM concentration of FA. The detection limit was found to be 0.74mM. Obviously, it is necessary to perform a directed genetic engineering of the yeast cells to improve their bioanalytical characteristics in the corresponding biosensors.
