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In this study, hydrolysates of large yellow croaker (Pseudosciaena crocea) roe were prepared by trypsin (Try‐pcRHs) and papain (Pap‐pcRHs). The degree of hydrolysis (DH), molecular weight distribution and iron chelating capacity of hydrolysates were investigated. Moreover, the chelation mechanism of iron‐hydrolysates complex also was investigated by amino acid composition analysis, fourier transform infrared spectrophotometer, fluorescence spectrophotometer and ultraviolet–visible spectroscopy. These results showed that Try‐pcRHs exhibited higher iron chelating ability with the EC50 value of 0.14 mg/ml than Pap‐pcRHs, which may be due to the higher DH and content of peptide <1 KDa. Furthermore, the Asp, Glu, Ser, Pro, Ile, Phe and His in Try‐pcRHs might contribute to the chelation reaction, and the groups of ‐COOH, C=O, N‐H and C‐O were involved in chealtiong of Try‐pcRHs with iron. Therefore, Try‐pcRHs have the potential to be used as iron supplements in food industries. Practical Applications: Large yellow croaker (Pseudosciaena crocea) roes are the main by‐products in the large yellow croaker processing. Previous studies have found that protein isolates from P. crocea roes composed of vitellogenin B, vitellogenin C and vitellogenin contain a large number of serine residues. In this study, we prepared hydrolysates from large yellow croaker roes, and the results showed that trypsin‐hydrolyzed large yellow croaker roes exhibited great iron‐chelating ability and has the potential to be used as a safe and healthy iron supplements in the food industry. [ABSTRACT FROM AUTHOR]