Abstract: To screen Lactobacilli strains that exhibit efficient purine nucleoside degradation capability and demonstrate probiotic potential, this study screened target strains from 88 lactic acid bacteria (LAB) strains using nucleoside degradation ability as an indicator. Consequently, eight strains with nucleoside degradation rates exceeding 80% were isolated. Further evaluations of their tolerance to simulated intestinal fluid and adhesion to Caco-2 cells led to the selection of strain CM33. This strain demonstrated exceptional nucleoside degradation performance, with inosine and guanosine degradation rates of 99.83% and 99.92%. Its tolerance to simulated intestinal fluid reached 96.82%, and its adhesion rate to Caco-2 cells was 61.36%. Identification via 16S-rDNA sequencing confirmed that CM33 belonged to the species Limosilactobacillus fermentum. However, strain CM33 exhibited limited tolerance to environments with pH levels of 2.5 and 3.0. This research provides a theoretical foundation for the development of LAB preparations with high purine nucleoside degradation capability.