High levels of soybean meal (SBM) in aquafeed leads to detrimental inflammatory response and oxidative stress in fish. In the present study, fermentation with Aspergillus awamori was conducted to explore the potential effects on improving the nutritional quality of soybean meal and the health status of turbot. A 63-day feeding trial (initial weight 8.53 ± 0.11 g) was carried out to evaluate the utilization of fermented soybean meal (FSM) by juvenile turbot. 0% (FM, control), 30% (S30, F30), 45% (S45, F45), and 60% (S60, F60) of fish meal were replaced with SBM or FSM, respectively. As the results showed, fermentation significantly reduced the contents of anti-nutritional factors in SBM, including raffinose (−98.8%), glycinin (−98.5%), β-conglycinin (−97.4%), trypsin inhibitors (−80%) and stachyose (−80%). A depression of fish growth performance and activities of superoxide dismutase and lysozyme were observed in S45 and S60 groups, while these inferiorities were only observed in F60 group. Meanwhile, fermentation also improved the heights of enterocytes and microvillus significantly in the F45 and F60 groups compared with those in SBM. An induced expression of anti-inflammatory cytokine transforming growth factor-β and depression of pro-inflammatory cytokines tumor necrosis factor-α and interleukin-1β in the distal intestine were observed in the F45 and F60 groups. Taken together, this study indicated that fermentation with Aspergillus awamori could improve the replacement level with soybean meal from 30% to 45% in turbot. • Aspergillus awamori fermentation reduced the contents of ANFs and improved the nutrition quality of soybean meal. • FSM enhanced antioxidant capacity, non-specific immunity and alleviated gut inflammatory response in turbot. • FSM could replace 45% of fishmeal protein in diets of juvenile turbot without adverse effects. [ABSTRACT FROM AUTHOR]