Efficient assay for respiration inhibitor using Saccharomyces cerevisiae

Saccharomyces cerevisiae를 이용한 효율적인 호흡저해제 검정법

  • Choi, Gyung-Ja (Screening Division, Korea Research Institute of Chemical Technology) ;
  • Kim, Jin-Cheol (Screening Division, Korea Research Institute of Chemical Technology) ;
  • Kim, Heung-Tae (Screening Division, Korea Research Institute of Chemical Technology) ;
  • Cho, Kwang-Yun (Screening Division, Korea Research Institute of Chemical Technology)
  • 최경자 (한국화학연구소 스크리닝연구부) ;
  • 김진철 (한국화학연구소 스크리닝연구부) ;
  • 김흥태 (한국화학연구소 스크리닝연구부) ;
  • 조광연 (한국화학연구소 스크리닝연구부)
  • Published : 2000.09.30

Abstract

A rapid assay to determine respiration inhibition of Saccharomyces cerevisiae by chemicals was developed. S. cerevisiae was harvested with two different liquid media, yeast extract-peptone-dextrose (YPD) medium capable of occurring both glucose fermentation and mitochondrial respiration, and non-fermentable carbon-yeast extract (NFY) medium capable of occurring respiration only Wells in 96-well plate were loaded with each cell suspension and various concentrations of 46 fungicides with various modes of action. n NFY medium, the non-fermentable carbon source, ethanol (NFY-E medium), glycerol (NFY-G medium) or lactate (NFY-L medium), was used. After incubation for $1{\sim}3$ days, minimum inhibitory concentrations (MICs) of the chemicals were recorded in the media. Of the 46 inhibitors employed in this study, four inhibitors of fungal respiration by blockage of electron flux in the mitochondrial respiratory chain, azoxystrobin, kresoxim-methyl, metominostrobin, and trifloxystrobin, exhibited strong antifungal activity in all of NFY media, but no activity in YPD medium. In contrast to this, five N-trihalomethylthio fungicides showed much stronger antifungal activities in YPD medium than three NFY media. Eleven fungicides inhibited growth of S. cerevisiae in all media and the other 26 fungicides showed no antifungal activity in all media. Thus, our rapid and efficient in vitro method can be considered as an alternative assay system for respiration inhibitor.