Menadione-catalyzed luminol luminescent assay as a novel evaluation method of ethanol tolerance in yeast cells.
In this study, ethanol inhibited the growth and glucose-induced proton release of yeast cells in a dose-dependent manner. On the other hand, ethanol tolerance of menadione-catalyzed luminol luminescence by yeast cells increased with increasing ethanol concentrations in the growth medium. The intracellular reduced-form nicotinamide adenine dinucleotide (NADH) concentration also increased with increasing ethanol concentrations in the medium and was enough to maintain constant menadione-catalyzed luminol luminescence. These facts suggest that the menadione-catalyzed luminol luminescent assay depending on a NADH:quinone reductase and NADH generation system is useful as a new evaluation assay for assessing the vitality of ethanol-stressed yeast cells, whereas the glucose-induced proton release assay is expected to be useful for the evaluation of cell growth under ethanol stress.
Yamashoji
Shiro
S
Shizuoka Institute of Science and Technology, Fukuroi, Shizuoka 437-8555, Japan. yamasho@ms.sist.ac.jp
D002384:Catalysis; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000737:chemistry / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol / Q000737:chemistry*; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D011522:Protons; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D024483:Vitamin K 3 / Q000737:chemistry*
D003470:Culture Media; D011522:Protons; D009243:NAD; D000431:Ethanol; D008165:Luminol; D024483:Vitamin K 3; D005947:Glucose