The aim of this study was to isolate and identify lactic acid bacteria (LAB) from a traditional fermented beverage, Boza, and to conduct an in-depth study on their fermentation and probiotic properties. The fermentation (acid production rate, acid tolerance, salt tolerance, amino acid decarboxylase activity) and probiotic properties (gastrointestinal tolerance, bile salt tolerance, hydrophobicity, self-aggregation, drug resistance, bacteriostatic properties) of the 16 isolated LAB were systematically analyzed by morphological, physiological, and biochemical tests and 16S rDNA molecular biology. This analysis utilized principal component analysis (PCA) to comprehensively evaluate the biological properties of the strains. The identified LAB included (9 strains), (2 strains), (2 strains), and (3 strains). These strains showed strong environmental adaptation at different pH (3.5) and temperature (45 °C), with different gastrointestinal colonization, tolerance, and antioxidant properties. All the strains did not show hemolytic activity and were inhibitory to , and showed resistance to kanamycin, gentamicin, vancomycin, and streptomycin. Based on the integrated scoring of biological properties by principal component analysis, S4 and S6 and S5 had excellent fermentation properties and tolerance and could be used as potential functional microbial resources.