Preparation, characterization, stability and replenishing calcium ability of Moringa oleifera leaf peptide-calcium...

Calcium deficiency has garnered significant attention as a global public health issue. A new generation of calcium supplements, peptide-calcium chelates, is expected to increase in market value. In this study, we produced MORP (MW < 1 kDa) from Moringa oleifera leaf protein via enzymatic hydrolysis for chelation with Ca²⁺ to produce MORP-Ca. SEM, EDS, FTIR and FS characterized the structure of MORP-Ca. The results indicate alterations in both the appearance and internal structure of MORP following calcium chelation. The functional groups of N-H, C-H, C-N, -C = O, -COO-, C-O, and -OH in MORP are involved in chelating Ca²⁺ to form MORP-Ca. In addition, MORP-Ca exhibits poor stability in the stomach; however, it demonstrates high stability in the intestine and under various temperature conditions. The results of the cellular experiments demonstrated that MORP-Ca is an effective promoter of calcium transport and absorption. MORP-Ca effectively increased bone mineral density and improved bone formation in animal studies. In addition, MORP-Ca supplementation improved the gut microbiota imbalance in rats fed a calcium-deficient diet, resulting in an increase in Firmicutes and a decrease in Actinobacteria. Thus, there is a connection between altered gastrointestinal flora and calcium absorption. LC-MS/MS and molecular docking analyses identified ARNEGRDL, RELIIGDR, YTPDYETK, YYTPDYETK, and IKFEFPAVDTL as key peptide sequences for the calcium-supplementing role of MORP (MW < 1 kDa). These results establish a theoretical foundation for the use of MORP-Ca as a calcium supplement or functional food.