Abstract:[Objective] To study the molecular mechanisms underlying improved acetic acid stress tolerance of the flocculating yeast Saccharomyces cerevisiae SPSC01 by zinc sulfate supplementation. [Methods] Global gene expression profiling was studied by comparative transcriptomic analysis using yeast cells of the log phase cells that were grown with or without 0.03 g/L zinc sulfate addition in the presence of 10.0 g/L acetic acid. [Results] Transcription levels of 50 genes were up-regulated and 162 genes were down-regulated when zinc sulfate was added in the fermentation medium. Genes involved in carbohydrate metabolism, methionine synthesis, and vitamin biosynthesis were up-regulated. In addition, genes encoding the antioxidant enzymes and other stress responsive genes were also up-regulated. [Conclusion] Zinc sulfate addition affected global gene transcription of S. cerevisiae, exerting positive effects on the expression of antioxidant enzymes, stress tolerance and redox balance, as well as energy metabolism. Our results indicate that zinc sulfate supplementation improves acetic acid tolerance of S. cerevisiae by regulating multiple genes and metabolic pathways.