Abstract In the present study the enzymatic production of orange seed protein hydrolysate (OSPH) from orange (Siavaraze, Citrus sinensis) seed protein using alcalase and pepsin enzymes under optimal conditions was evaluated. Then, the effect of OSPH type and chitosan concentration (0, 0.1, 0.2, and 0.4% w/v) on physicochemical characteristics, release rate, stability, antioxidant activity, and morphology of the loaded nanoliposomes was assessed. The results revealed that the particle size, polydispersity index (PDI), and encapsulation efficiency (EE) were influenced by the type of loaded OSPH and chitosan concentration. The zeta potential value in the nanoliposomes after loading with OSPH obtained from alcalase and pepsin changed from −8.87 mV to −9.45 mV and +4.41 mV, respectively. In addition, this index shifted toward positive values after coating with chitosan. The freeze-thaw stress and storing at 4 °C (for 4 weeks) led to the highest (about 10-fold) and lowest (about 2-fold) changes in the nanoparticles size, respectively. Among different treatments, the nanoliposome coated by 0.2% W/V chitosan showed decreased peptide release rate, retention of EE, and physical stability under different conditions. Evaluation of the chemical structure using FTIR indicated the positioning of the peptides inside the polar regions, bilayer-membrane, and their ionic complexation with liposome phosphate groups. Moreover, the evaluation of the nanoparticles’ morphology indicated spherical particles with smooth surfaces, confirming the dynamic light scattering (DLS) results. The results of this study indicate the efficacy of coated liposomes in protection, control of release and maintaining the antioxidant activity of peptides.

中文翻译：

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橙籽蛋白水解物负载壳聚糖纳米脂质体的理化性质

摘要 在本研究中，评估了在最佳条件下使用碱性蛋白酶和胃蛋白酶从橙子（Siavaraze，Citrus sinensis）种子蛋白中酶促生产橙子种子蛋白水解物 (OSPH)。然后，评估了 OSPH 类型和壳聚糖浓度（0、0.1、0.2 和 0.4% w/v）对负载纳米脂质体的理化特性、释放速率、稳定性、抗氧化活性和形态的影响。结果表明，粒径、多分散指数 (PDI) 和包封效率 (EE) 受负载 OSPH 类型和壳聚糖浓度的影响。加载从碱性蛋白酶和胃蛋白酶获得的 OSPH 后，纳米脂质体中的 zeta 电位值分别从 -8.87 mV 变为 -9.45 mV 和 +4.41 mV。此外，用壳聚糖包衣后，该指数向正值移动。冻融压力和在 4 °C 下储存（4 周）分别导致纳米颗粒尺寸的最高（约 10 倍）和最低（约 2 倍）变化。在不同处理中，0.2% W/V壳聚糖包覆的纳米脂质体在不同条件下的肽释放率、EE保留率和物理稳定性均降低。使用 FTIR 对化学结构的评估表明肽在极性区域、双层膜内的定位以及它们与脂质体磷酸基团的离子络合。此外，对纳米颗粒形态的评估表明具有光滑表面的球形颗粒，证实了动态光散射 (DLS) 结果。