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Alginate-based fibers with good biocompatibility and non-immunogenicity have wide applications in biomedical materials. However, low strength and poor water resistance still limit their application in tissue engineering materials. In this study, a novel alginate ester/Antarctic krill protein (AE/AKP) composite fiber with a "covalent crosslinking and hydrogen bonding" structure was synthesized based on acid-catalyzed phase separation reaction spinning by using sodium alginate as the matrix, AKP as a hydrogen bond acceptor, and polyol as the crosslinking agent. The mechanism of hydrogen bond interaction and its relationship with the fiber structure and properties were investigated by Fourier transform infrared spectroscopy and X-ray diffraction. The tensile strength of the fiber is 4.29 cN/dtex, which is 10.28% higher than that of the AE fiber and 96.79% higher than that of the cross-linked ion fiber, and the crystallinity reaches 30.57%. Moreover, the cytotoxicity test results showed that the fiber was non-toxic. These results indicate that AE/AKP composite fibers have great potential as tissue engineering materials. [ABSTRACT FROM AUTHOR]