title: Strengths and weaknesses in the determination of Saccharomyces cerevisiae cell viability by ATP-based bioluminescence assay.
abstract: Due to its sensitivity and speed of execution, detection of ATP by luciferin-luciferase reaction is a widely spread system to highlight cell viability. The paper describes the methodology followed to successfully run the assay in the presence of yeast cells of two strains of the yeast Saccharomyces cerevisiae, BY4741 and CEN.PK2-1C and emphasizes the importance of correctly determining the contact time between the lysing agent and the yeast cells. Once this was established, luciferin-luciferase reaction was exploited to determine the maximum specific rate of growth, as well as cell viability in a series of routine tests. The results obtained in this preliminary study highlighted that using luciferin-luciferase can imply an over-estimation of maximum specific growth rate with respect to that determined by optical density and/or viable count. On the contrary, the bioluminescence assay gave the possibility to highlight, if employed together with viable count, physiological changes occurring in yeast cells as response to stressful environmental conditions such as those deriving from exposure of yeast cells to high temperature or those depending on the operative conditions applied during fed-batch operations.
authors: Department of Industrial Engineering, Università di Salerno, Via Ponte Don Melillo, 84084 Fisciano, Salerno, Italy.
authors: Lucia
authors: L
authors: Paciello
authors: Francesco Cristino
authors: FC
authors: Falco
authors: Carmine
authors: C
authors: Landi
authors: Palma
authors: P
authors: Parascandola
mesh_terms: D000255:Adenosine Triphosphate; D061249:Batch Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D018533:Biomass; D019149:Bioreactors; D002473:Cell Wall / Q000187:drug effects; D015169:Colony Count, Microbial; D003720:Densitometry; D049454:Firefly Luciferin; D006358:Hot Temperature; D049409:Luciferases, Firefly; D008163:Luminescent Measurements; D009172:Mycology / Q000379:methods; D009391:Nephelometry and Turbidimetry; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000187:drug effects / Q000254:growth & development; D013045:Species Specificity; D013053:Spectrophotometry; D013997:Time Factors
chemical_list: D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D049409:Luciferases, Firefly