Amperometric monitoring of redox activity in intact, permeabilised and lyophilised cells of the yeast Hansenula polymorpha

An effect of permeabilisation and lyophilisation of the yeast cells Hansenula polymorpha on their electrochemical behaviour in the presence of mediators, substrates (formaldehyde, glucose, methanol, ethanol), and cofactors (NAD+, NADP+, NADH, NADPH, glutathione) has been studied. Two amperometric techniques differing in the cell immobilisation methods were applied. The cells of a wild strain (356) and mutant strains (C-105 and KCA 33) of the yeast, grown in the presence of glucose or methanol, were used in the experiments. The intact cells revealed the highest reduction rates of mediators, 2,6-dichlorphenolindophenol (DCIP) and 2,4-benzoquinone (BQ), as measured by amperometry. The addition of formaldehyde significantly enhanced the response, if the cells were grown in the presence of glucose. The permeabilised cells showed the lowest current level in the presence of DCIP and BQ and no response to the addition of formaldehyde and NAD+. However, the addition of NADH gave significant current surge. All these phenomena imply that the permeabilised cells lost cofactors and the activity of dehydrogenases producing NADH, but they remained the activity of NADH-ubiquinone oxidoreductase and of some components of the electron transport chain. The electrochemical behaviour of the lyophilised cells shows they are heterogeneous. The partial degradation of the outer membrane of the cells after their lyophilisation was electrochemically confirmed.