Succinic acid (SA) is a valuable chemical with broad applications
however, its high concentrations can inhibit yeast cells, reducing fermentation efficiency. In this study, adaptive laboratory evolution was used to enhance yeast tolerance to SA, resulting in several strains capable of growing in medium with 40 g/L SA. Subsequently, whole genome sequencing of the evolved strains was conducted to identify beneficial genetic adaptations. A total of eleven gene mutations were identified across three independent evolutionary lineages, six of which are associated with cell wall functionality and contribute to SA tolerance. Specifically, the deletion of MNN4 impairs mannose side chains and significantly increases resistance to SA. Additionally, the GAS1