Proteins related to antioxidant defence systems and energy maintenance play a vital role in resisting starvation in fish. However, studies on the underlying molecular mechanism of starvation stress in the jade perch Scortum barcoo are very limited. Thus, the proteomic changes in S. barcoo during fasting were investigated by stable isotope labelling and high‐performance liquid chromatography‐tandem mass spectrometry (iTRAQ‐LC‐ MS/MS) in the present study. Therefore, a total of 3324 proteins were identified, including 134 that were significantly altered (p < 0.05, fold change >1.20 or <0.83) in the experimental groups compared with the control group. Among all the differentially expressed proteins, only protein‐disulphide reductase (TXNDC12) was upregulated. One‐third of the differentially expressed proteins were enriched in the amino acid metabolism pathway, tricarboxylic acid (TCA) cycle pathway, fatty acid metabolism pathway and oxidative phosphorylation pathway. Proteins differentially expressed in the experimental group, including AASS, OGDH, ACAD8, FAH, BHMT, AOC3, AOC2 and HIBADH, are components of pathways that control the metabolism of amino acids. The results suggested the involvement of antioxidant defence in the response to an oxidative imbalance, and GSH, SOD and AKP were regulated under conditions of starvation stress. In conclusion, the data indicated that starvation‐induced changes in protein levels that might affect metabolism and the antioxidant system.