Separation and Identification of Novel Antithrombotic Protein and Peptides from Shrimp Paste Hydrolysates

shrimp paste via enzymatic-assisted fermentation to enhance its dual antithrombotic profile and identify the responsible components. Optimal activity was achieved by using 1% chitosanase at 45 °C and pH 7.0 for 6 h, followed by fermentation at 25 °C for 15 d. A fibrinolytic fragment (approximately 11.36 kDa) from the SIT-AKD region of protein A0A423T5F8 showed activity equivalent to 40.28 μg urokinase at 21 μg. Meanwhile, peptides PSEPSKPVTCKPR and GIAEGCDYPWR exhibited 1.35-fold and 1.19-fold higher anticoagulant activity than heparin sodium at 3 mg/mL. Molecular docking indicated that hydrogen-bonding interactions with Trp60, Glu192, Gly216, and Arg221 underlie the anticoagulant mechanism. These findings support the high-value utilization of shrimp paste.