The present research examined the impact of L-glutamic acid (Glu) supplementation on the growth performance, muscle composition, gene expression correlated with muscle growth, and intestinal health of largemouth bass. There were 525 fish in total, which were distributed randomly into five groups. Each group had three replicates, and each replicate consisted of 35 fish. Groups with control and experimental diets were assigned glutamic acid amounts of 0.2%, 0.4%, 0.6%, and 0.8%. The findings demonstrated that glutamic acid supplementation enhanced growth performance, feed intake (FI), and condition factor (CF), with the best value being attained at 0.4% Glu. The mean muscle fiber area was increased and the muscle fiber density was decreased in the 0.6% Glu group. The levels of total amino acids and specific amino acids, such as glutamic acid, aspartic acid, leucine, valine, alanine, and glycine, were shown to be higher in the 0.6% Glu group. In the 0.6% Glu group, the mRNA expression levels of atrogin-1, murf-1, foxo3a, and 4e-bp1 were decreased compared to the control group. Conversely, the mRNA expression levels of myf5, myog, myod, s6k1, tor, akt, and pi3k were increased in the 0.6% Glu group compared to the control group. The 0.4% Glu group had higher intestinal amylase, lipase, and protease activities and greater villus height, villus width, and muscle thickness. In summary, Glu can support largemouth bass growth, muscular development, intestinal digestion, and absorption.