Novel calcium chelating peptide as well as preparation method and application thereof
A new type of calcium chelation technology, applied in the field of new calcium chelating peptides and their preparation, to achieve the effect of improving absorption and bioavailability, simple operation, and promoting calcium transport
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Embodiment 1
[0053] This embodiment provides a calcium chelating peptide whose amino acid sequence is: YQEPVIAPKL().
[0054] The calcium chelating peptide can be prepared by the following preparation method:
[0055] (1) Extraction of tilapia bone collagen
[0056] (1) Cleaning of tilapia skeleton raw materials
[0057] After the fish skeleton is thawed, impurities are removed and cleaned with tap water, and finally the fish skeleton is decapitated.
[0058] (2) De-meat treatment of tilapia skeleton
[0059] Cut the fish skeleton into about 5 cm, take 1 kg of raw material, add 5 g of neutral protease and 1000 mL of water, and carry out enzymatic hydrolysis on the fish skeleton for 2 hours (55°C, pH 7.5) to remove the minced meat attached to the bone. Impurities are washed away after enzymatic hydrolysis.
[0060] (3) Alkaline Elution Degreasing of Tilapia Bones
[0061] Put the fish bones into 0.1mol / L NaOH solution (w / v1:10) and stir for 24h, and wash until neutral.
[0062] (4) De...
Embodiment 2
[0131] In this example, the calcium chelating ability of the calcium chelating peptide and its effect on the calcium transport of the Caco-2 cell monolayer were studied, and the specific process was as follows.
[0132] (1) Peptide synthesis
[0133] Synthetic peptides (YQEPVIAPKL) with a purity higher than 95% were obtained from Synpeptide Co., Ltd (Nanjing, China), and their molecular weight was determined by LC / ESI-MS, which was 1157.627 Da.
[0134] Table 4 shows the homologous proteins of calcium chelating peptides.
[0135] Homologous proteins of table 4 calcium-chelating peptides
[0136]
[0137] The calcium sequestration activity of the synthetic peptides was determined by using the o-cresolphthalein colorimetric method. The calcium chelating activity of YQEPVIAPKL was 28.4 mg / g, indicating that the shorter peptide YQEPVIAPKL has more calcium chelating sites.
[0138] (2) Cell culture
[0139] Caco-2 cells were purchased from American Type Culture Collection (R...
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