Coix seed (Coix lachryma-jobi L.) is an important nourishing food and traditional Chinese medicine. The role of their bioactive constituents in physiology and pharmacology has received considerable scientific attention. However, very little is known about the role of coix seed bioactive components in the growth of Limosilactobacillus reuteri (L. reuteri). This study aimed to evaluate the effects of coix seed extract (CSE) on the growth, acidifying activity, and metabolism of L. reuteri. The results showed that CSE can increase the growth and acidifying activity of L. reuteri compared with the control group. During the stationary phase, the viable bacteria in the medium supplemented with coix seed oil (CSO, 13.72 Log10 CFU/mL), coix polysaccharide (CPO, 12.24 Log10 CFU/mL), and coix protein (CPR, 11.91 Log10 CFU/mL) were significantly higher (p < 0.05) than the control group (MRS, 9.16 Log10 CFU/mL). CSE also enhanced the biosynthesis of lactic acid and acetic acid of L. reuteri. Untargeted metabolomics results indicated that the carbohydrate metabolism, amino acid metabolism, and nucleotide metabolism activities of L. reuteri were increased after adding CSE. Furthermore, CSE increased the accumulation of bioactive metabolites, such as phenyl lactic acid, vitamins, and biotin. Overall, CSE may have prebiotic potential and can be used to culture L. reuteri with high viable bacteria.