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In recent years, short-term composting techniques have been widely applied in oyster mushroom cultivation, but there is still a lack of systematic research on their impact on the nutritional and functional properties of fruiting bodies. In this study, the microbial inoculant Streptomyces thermoviolaceus BUA-FM01 (ST) was applied in the short-term composting process for oyster mushroom cultivation. The agronomic traits, nutritional composition, flavor compounds, and antioxidant activity of fruiting bodies from the first three flushes were evaluated. The results show that microbial inoculation significantly (p < 0.05) reduced the total carbon content and C/N ratio of the composted substrates and, furthermore, increased the total yield of the fruiting bodies. Moreover, microbial inoculation significantly (p < 0.05) increased the crude protein, crude polysaccharide, total amino acid, and essential amino acid contents of the fruiting bodies. The fruiting bodies of the first flush of ST treatment possessed the highest umami amino acid content and equivalent umami concentration value. Furthermore, microbial inoculation significantly (p < 0.05) enhanced the scavenging ability of crude polysaccharides toward free radicals. The results indicate that microbial inoculation has many benefits for the composting cultivating process of oyster mushrooms and good application prospects. [ABSTRACT FROM AUTHOR]