The purification, identification and bioactivity study of a novel calcium-binding peptide from casein hydrolysate

Liao, Wanwen; Liu, Shuojun; Liu, Xiaorong; Duan, Shengwen; Xiao, Suyao; Yang, Zhennai; Cao, Yong; Miao, Jianyin

doi:10.1039/c9fo01383k

Cited by 30 publications

(17 citation statements)

References 51 publications

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“…Zhang identified the novel calcium-binding peptide KGDPGLSPGK from Pacific cod bone gelatin hydrolysate and verified that Asp-3 and Lys-10 are calcium-binding sites. A novel calcium-binding peptide, VLPVPQK, identified from casein, effectively promotes the transport and absorption of calcium ions, and also contains Val, Leu, Pro, and Lys [ 55 ]. Calcium-binding peptides extracted from bovine serum have also been confirmed to contain Lys.…”

Section: Resultsmentioning

confidence: 99%

Isolation, Purification and Structure Identification of a Calcium-Binding Peptide from Sheep Bone Protein Hydrolysate

Wang

Sun

et al. 2022

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To isolate a novel peptide with calcium-binding capacity, sheep bone protein was hydrolyzed sequentially using a dual-enzyme system (alcalase treatment following neutrase treatment) and investigated for its characteristics, separation, purification, and structure. The sheep bone protein hydrolysate (SBPH) was enriched in key amino acids such as Gly, Arg, Pro, Leu, Lys, Glu, Val, and Asp. The fluorescence spectra, circular dichroism spectra, and Fourier-transform infrared spectroscopy results showed that adding calcium ions decreased the α-helix and β-sheet content but significantly increased the random and β-turn content (p < 0.05). Carboxyl oxygen and amino nitrogen atoms of SBPH may participate in peptide–calcium binding. Scanning electron microscopy and energy dispersive spectrometry results showed that SBPH had strong calcium-chelating ability and that the peptide–calcium complex (SBPH–Ca) combined with calcium to form a spherical cluster structure. SBPH was separated and purified gradually by ultrafiltration, gel filtration chromatography, and reversed-phase high-performance liquid chromatography. Liquid chromatography-electrospray ionization/mass spectrometry identified the amino acid sequences as GPSGLPGERG (925.46 Da) and GAPGKDGVRG (912.48 Da), with calcium-binding capacities of 89.76 ± 0.19% and 88.26 ± 0.25%, respectively. The results of this study provide a scientific basis for the preparation of a new type of calcium supplement and high-value utilization of sheep bone.

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Section: Resultsmentioning

confidence: 99%

Isolation, Purification and Structure Identification of a Calcium-Binding Peptide from Sheep Bone Protein Hydrolysate

Wang

Sun

et al. 2022

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“…For example, EPQWFL derived from microalgae, VLPVPQK derived from milk, and RVYFFKGKQYWE derived from vitronectin have been shown to have superior osteogenic activity. 35–39 In addition, peptides containing glutamine sequences have been identified in some endogenous osteogenic proteins, such as GQGFSYPYKAVFSTQ derived from BMP and ALKRQGRTLYGFGG from OPG. 40 In this study, the content of glutamic acid/glutamine in SCIP was 21.1%, which is close to the recognized high-quality proteins of wheat and corn.…”

Section: Discussionmentioning

confidence: 99%

Novel peptides from sea cucumber intestinal hydrolysates promote longitudinal bone growth in adolescent mice through accelerating cell cycle progress by regulating glutamine metabolism

et al. 2022

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“…Calcium chelating activity was determined by the o-cresol phthalein colorimetry method ( 15 ). Five milligrams of CPPs freeze-dried powder were mixed with 1 mL of 5 mM CaCl 2 solution and 2 mL of phosphate buffer.…”

Section: Methodsmentioning

confidence: 99%

“…For example, Liu et al found that CPPs can promote the transport of calcium in Caco-2 monolayers ( 14 ). Liao et al purified and identified a novel calcium-binding peptide with good calcium transport capacity from casein hydrolysate ( 15 ). Studies have shown that CPPs have good osteogenic activity.…”

Section: Introductionmentioning

confidence: 99%

Preparation, characterization, and osteogenic activity mechanism of casein phosphopeptide-calcium chelate

et al. 2022

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Casein phosphopeptides (CPPs) are good at calcium-binding and intestinal calcium absorption, but there are few studies on the osteogenic activity of CPPs. In this study, the preparation of casein phosphopeptide calcium chelate (CPP-Ca) was optimized on the basis of previous studies, and its peptide-calcium chelating activity was characterized. Subsequently, the effects of CPP-Ca on the proliferation, differentiation, and mineralization of MC3T3-E1 cells were studied, and the differentiation mechanism of CPP-Ca on MC3T3-E1 cells was further elucidated by RNA sequencing (RNA-seq). The results showed that the calcium chelation rate of CPPs was 23.37%, and the calcium content of CPP-Ca reached 2.64 × 105 mg/kg. The test results of Ultraviolet–Visible absorption spectroscopy (UV) and Fourier transform infrared spectroscopy (FTIR) indicated that carboxyl oxygen and amino nitrogen atoms of CPPs might be chelated with calcium during the chelation. Compared with the control group, the proliferation of MC3T3-E1 cells treated with 250 μg/mL of CPP-Ca increased by 21.65%, 26.43%, and 28.43% at 24, 48, and 72 h, respectively, and the alkaline phosphatase (ALP) activity and mineralized calcium nodules of MC3T3-E1 cells were notably increased by 55% and 72%. RNA-seq results showed that 321 differentially expressed genes (DEGs) were found in MC3T3-E1 cells treated with CPP-Ca, including 121 upregulated and 200 downregulated genes. Gene ontology (GO) revealed that the DEGs mainly played important roles in the regulation of cellular components. The enrichment of the Kyoto Encyclopedia of Genes and Genomes Database (KEGG) pathway indicated that the AMPK, PI3K-Akt, MAPK, and Wnt signaling pathways were involved in the differentiation of MC3T3-E1 cells. The results of a quantitative real-time PCR (qRT-PCR) showed that compared with the blank control group, the mRNA expressions of Apolipoprotein D (APOD), Osteoglycin (OGN), and Insulin-like growth factor (IGF1) were significantly increased by 2.6, 2.0 and 3.0 times, respectively, while the mRNA levels of NOTUM, WIF1, and LRP4 notably decreased to 2.3, 2.1, and 4.2 times, respectively, which were consistent both in GO functional and KEGG enrichment pathway analysis. This study provided a theoretical basis for CPP-Ca as a nutritional additive in the treatment and prevention of osteoporosis.

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The purification, identification and bioactivity study of a novel calcium-binding peptide from casein hydrolysate

Abstract: In this study, a novel calcium-binding peptide from casein hydrolysate was purified using reversed-phase high performance liquid chromatography and sequenced by high-performance liquid chromatography-mass spectrometry (MS)/MS.

Cited by 30 publications

References 51 publications

Isolation, Purification and Structure Identification of a Calcium-Binding Peptide from Sheep Bone Protein Hydrolysate

Isolation, Purification and Structure Identification of a Calcium-Binding Peptide from Sheep Bone Protein Hydrolysate

Novel peptides from sea cucumber intestinal hydrolysates promote longitudinal bone growth in adolescent mice through accelerating cell cycle progress by regulating glutamine metabolism

Preparation, characterization, and osteogenic activity mechanism of casein phosphopeptide-calcium chelate

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