Our previous work indicated that the quorum sensing (QS) effect could regulate the oxidative tolerance of Saccharomyces cerevisiae, and QS may impact oxidative and antioxidative metabolisms of S. cerevisiae by regulating the RCK1 gene. Therefore, this work proposed a reasonable logic that RCK1 could play roles in regulating the oxidative and antioxidative metabolisms of yeast cells. The results presented here suggested that the overexpression of RCK1 has a regulatory effect on the reduction of ROS level and the promotion of oxidative tolerance of S. cerevisiae. The overexpression of RCK1 promoted the ROS generation through activating the MAPK pathway; on the other hand, RCK1-regulated antioxidative metabolism played a more significant role to realize lower ROS level and higher oxidative tolerance of S288c-RCK1 and ΔARO80-RCK1 strains. To improve the fermentation performance of yeast while circumventing metabolic burden, a recombinant strain with over time-controlled overexpression of the RCK1 gene (i.e., S288c'-RCK1 strain) derived from S288c strain was successfully constructed to achieve artificial regulation of yeast oxidative tolerance. Transcriptomics analysis was further performed on both S. cerevisiae wild-type and S288c'-RCK1 strains to identify differentially expressed genes and analyze their functional pathway classification. This work is instructive for artificially modulating the oxidative tolerance of strains to enhance the fermentation performance of yeast.