To screen lactic acid bacteria (LAB) with high ethanol tolerance and degradation capacity and to elucidate their underlying metabolic regulatory mechanisms. More than 140 LAB strains were isolated from various food sources. Ethanol tolerance was evaluated based on their growth in media containing 8% and 12% (φ) ethanol, and ethanol degradation capacity was further determined under 4%, 8%, and 12% (φ) ethanol conditions. In addition, real-time quantitative PCR was employed to analyze the expression levels of key functional genes in the selected strain under ethanol stress. Six LAB strains exhibited good growth at 12% (φ) ethanol, among which Lactiplantibacillus plantarum YZ-010 showed the strongest ethanol tolerance and degradation ability, with a degradation rate of 33.01 wt.% at 4% (φ) ethanol. Under ethanol stress, genes related to alcohol and aldehyde dehydrogenases (adh1, aldH, Nox, and adh zin-binding) were significantly upregulated, promoting the conversion of ethanol into acetyl-CoA. Meanwhile, genes involved in glycerolipid metabolism (plsC and dhaT), aminoacyl-tRNA biosynthesis (thrS), peptidoglycan synthesis (dacA), and purine transport (NCS2) were upregulated, whereas amino acid metabolism-related genes (argG, argH, and gadB) were downregulated. L. plantarum YZ-010 exhibits strong ethanol tolerance and degradation capacity, which are associated with enhanced alcohol-aldehyde conversion, maintenance of cellular structural stability, and metabolic reprogramming. These findings provide a theoretical basis for the development of probiotic-based anti-alcohol functional products and for understanding the molecular mechanisms of ethanol tolerance in LAB.