405 nm and 450 nm Photoinactivation of Saccharomyces cerevisiae.

Abstract

Photoinactivation of bacteria with visible light has been reported in numerous studies. Radiation around 405 nm is absorbed by endogenous porphyrins and generates reactive oxygen species that destroy bacteria from within. Blue light in the spectral range of 450-470 nm also exhibits an antibacterial effect, but it is weaker than 405 nm radiation, and the photosensitizers involved have not been clarified yet, even though flavins and porphyrins are possible candidates. There are significantly fewer photoinactivation studies on fungi. To test if visible light can inactivate fungi and to elucidate the mechanisms involved, the model organism Saccharomyces cerevisiae (DSM no. 70449) was irradiated with violet (405 nm) and blue (450 nm) light. The mean irradiation doses required for a one log reduction of colony forming units for this strain were 182 J/cm² and 526 J/cm² for 405 nm and 450 nm irradiation, respectively. To investigate the cell damaging mechanisms, trypan blue staining was performed. However, even strongly irradiated cultures hardly showed any stained S. cerevisiae cells, indicating an intact cell membrane and thus arguing against the previously suspected mechanism of cell membrane damage during photoinactivation with visible light at least for the investigated strain. The results are compatible with photoinactivated Saccharomyces cerevisiae cells being in a viable but nonculturable state. To identify potential fungal photosensitizers, the absorption and fluorescence of Saccharomyces cerevisiae cell lysates were determined. The spectral absorption and fluorescence results are in favor of protoporphyrin IX as the most important photosensitizer at 405 nm radiation. For 450 nm irradiation, riboflavin and other flavins may be the main photosensitizer candidates, since porphyrins do not play a prominent role at this wavelength. No evidence of the involvement of other photosensitizers was found in the spectral data of this strain.