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A calcium-chelating peptide is considered to have the ability to improve calcium absorption. In this study, Pacific cod skin gelatin hydrolysates treated with trypsin for 120 min exhibited higher calcium-chelating activity. Sequential chromatography, involving hydroxyapatite affinity chromatography and reversed phase high performance liquid chromatography, was used for the purification of calcium-chelating peptides. Two novel peptides with the typical characteristics of collagen were sequenced as GDKGESGEAGER and GEKGEGGHR based on LC-HRMS/MS, which showed a high affinity to calcium. Calcium-peptide complexation was further characterized by ESI-MS (MS and MS/MS) and FTIR spectroscopy. The results showed that the complexation of the two peptides with calcium was conducted mainly at the ratio of 1 : 1. The amino terminal group and the peptide bond of the peptide backbone as well as the amino group of the lysine side chain and the carboxylate of the glutamate side chain were the possible calcium binding sites for the two peptides. Meanwhile, several amino acid side chain groups, including the hydroxyl (Ser) and carboxylate (Asp) of GDKGESGEAGER and the imine (His) of GEKGEGGHR, were crucial in the complexation. The arginine residue in GEKGEGGHR also participated in the calcium coordination. Additionally, several active fragments with calcium-chelating activity were obtained using MS/MS spectra, including GDKGESGEAGE, GEAGER, GEK, EKG and KGE. This study suggests that gelatin-derived peptides have the potential to be used as a calcium-chelating ingredient to combat calcium deficiency.

Highlights Ultrasound improves protein extraction rate by 16.56% than alkaline extraction. Ultrasound can affect iron-chelating peptide generation by protein extraction. Two alcalase hydrolysates own similar iron-chelating rate and binding sites. Main peptide sequences have different structural characteristic from each other.

The purpose of this study was to construct a glycogen (Gly)-based nanoparticle (NP) with liver-targeted and redox response to effectively deliver resveratrol (Res) for improving nonalcoholic fatty liver disease (NAFLD). Herein, Gly was modified using α-lipoic acid (α-LA) and lactobionic acid (Lac) to obtain an amphiphilic polymer (Gly−LA−Lac), which was self-assembled in water and then encapsulated in Res to form Res NPs with excellent stability. As expected, the Res NPs exhibited liver-targeted and redox response release behavior. In vitro cell studies demonstrated that the nanocarrier treatment enhanced the cellular uptake of Res and reduced oxidative stress and inflammatory factor levels. Meanwhile, the in vivo tests proved that the nanocarriers effectively reduced hepatic lipid accumulation and oxidative stress levels via regulating the TLR4/NF-κB signal pathway to improve liver damage in NAFLD mice. In conclusion, this study provides a promising strategy through the construction of Gly-based nanocarriers for the encapsulation of Res to effectively alleviate the process of NAFLD.