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This study aimed to explore the effects of glutamate (Glu) supplementation on the growth performance, carcass traits, meat quality, composition of amino acids and fatty acids in the longissimus dorsi muscle, and the colonic microbial community of Shaziling pigs. A total of 48 healthy male Shaziling pigs (150 d, 31.56 ± 0.95 kg) were randomly assigned to two groups, and fed a basal diet with no supplement (control group) or supplemented with 1% Glu (Glu group) for 51 d, and 6 pigs per group were finally slaughtered. Glu significantly increased the average daily gain (P= 0.039), lean percentage (P= 0.023), and intramuscular fat (IMF) content (P= 0.015), and decreased the fat percentage (P= 0.021) of Shaziling pigs. In the muscle, Glu increased the concentrations of inosine-5’-monophosphate (P= 0.094), Fe (P= 0.002), Cu (P= 0.052), and monounsaturated fatty acids (MUFAs) (P= 0.024), and decreased the content of C18:2n6 (P= 0.011), n6 polyunsaturated fatty acids (n6 PUFAs) (P= 0.014), and PUFAs (P= 0.014). Moreover, Glu significantly upregulated the mRNA expression of adipogenesis-related genes (FAS, SREBP-1C) (P= 0.032, P= 0.026) and muscle growth-related genes (MyHCⅡb, MyHCⅡx) (P= 0.038, P= 0.019) in the muscle, and increased the relative abundance of Spirochaetota (P< 0.001) and the acetic acid content in the colon (P= 0.039). Correlation analysis indicated that the acetic acid content was positively correlated with the relative Spirochaetota abundance and the IMF content, and a negative trend with the fat percentage of Shaziling pigs. In conclusion, these results indicated that Glu could simultaneously increase the lean percentage and IMF content and decrease the fat percentage of Shaziling pigs, and these beneficial effects may be related to increased colonic Spirochaetota abundance and acetic acid concentrations.