Lactobridge protein source calcium chelating peptide and application thereof

Abstract

The invention discloses a lactobridge protein source calcium chelating peptide and application thereof, and belongs to the technical field of biological small molecule active peptides. The calcium chelating peptide provided by the invention is safe and non-toxic, has better physical and chemical activity compared with a traditional calcium supplement, has the calcium chelating capacity of 419.36 mu g / mg or above, can promote calcium transport in a Caco-2 cell monolayer, can improve absorption and bioavailability of calcium in a human body, and is used as a calcium supplement preparation. According to the invention, a new protein source can be developed for calcium supplements, and an important contribution is made to high-value utilization of dairy products.

Description

Technical Field

[0001] The present invention relates to a milk bridge protein-derived calcium chelating peptide and its application, belonging to the technical field of bioactive small peptides. Background Art

[0002] Calcium is the main component of bone minerals. The human body mainly obtains calcium by ingesting food. It has been found through research that calcium metabolic imbalance caused by insufficient calcium intake is the cause of various diseases in people of all ages. At present, calcium deficiency has become a worldwide nutritional phenomenon.

[0003] Currently, there are mainly three types of calcium supplements on the market:

[0004] Inorganic calcium: The first-generation calcium preparations are mainly inorganic salts. Their disadvantages are low solubility and the formation of precipitates in the alkaline environment of intestinal juice, resulting in low soluble calcium content. In addition, this type of calcium preparation will stimulate the stomach after reacting with gastric acid and generate gas in the gastrointestinal tract, causing side effects such as flatulence.

[0005] Organic acid calcium: The second-generation calcium preparations are mainly organic acid salts. Compared with the first-generation calcium preparations, their solubility and absorption rate have been improved, but their calcium content is generally low.

[0006] Organic acid chelated calcium: The third-generation calcium preparations are mainly organic acid chelated calcium sources, mainly chelates formed by amino acids and calcium ions, and compound calcium preparations made with vitamin C and vitamin D as excipients. This generation of calcium preparations has good dissolution effects and can be absorbed without dissociating into calcium ions, so the calcium absorption rate is relatively high. However, its transport will be affected by food, so it is only suitable for taking calcium on an empty stomach.

[0007] In recent years, with the development of small peptides, the research and development process of the fourth-generation calcium preparations has been promoted. The fourth-generation calcium preparations refer to chelates formed by chelating small peptides and calcium ions with coordination covalent bonds. Compared with the third-generation calcium preparations, in terms of structure, due to the participation of non-chelated oxygen peptide bonds in the coordination effect, the peptide-calcium complex is more stable than amino acid chelated calcium. In terms of absorption, the amino and carboxyl groups at the amino acid terminal participate in the calcium chelation simultaneously, forming a double-ring chelation structure without free carboxyl groups and cannot be completely absorbed by itself. However, due to the steric hindrance of the peptide-calcium complex, there are free carboxyl groups and it is absorbed as a whole in the small intestine and releases calcium ions when needed. As a carrier, small bioactive peptides themselves have the characteristics of fast transport, low energy consumption, and not easy to saturate, and have the ability to prevent calcium phosphate precipitation, making the peptide-calcium complex always in a soluble calcium state and directly decomposed into an ionic state at the absorption site for absorption, thus greatly improving the calcium absorption rate.

[0008] Literature reports (Liao W, Liu S, Liu X, et al. The purification, identification and bioactivity study of a novel calcium-binding peptide from casein hydrolysate. Food Funct. 2019;10(12):7724-7732. doi:10.1039 / c9fo01383k) show that the heptapeptide VLPVPQK derived from casein hydrolysate has strong calcium-binding activity (129.46 mg / g) and can effectively enhance the transport and absorption of calcium in the Caco-2 cell monolayer in a concentration-dependent manner.

[0009] Lactopontin (LPN) is a highly phosphorylated protein isolated from mammalian milk. LPN and its hydrolysates are rich in acidic amino acids and phosphoserine residues, which can bind a large amount of calcium ions with strong affinity. However, its binding sites and mechanism of action are still unclear, which limits its application as a calcium supplement product. LPN can be enzymatically hydrolyzed into small peptides, thereby exposing the active center.

[0010] Therefore, the isolation and preparation of calcium-chelating peptides from LPN hydrolysates is of great significance for the basic theoretical research on the regulation of biomineralization by phosphorylated proteins, can develop new protein sources for calcium supplements, and make important contributions to the high-value utilization of dairy products. Summary of the Invention

[0011] To solve the above problems, the present invention provides a calcium-chelating peptide DDLDDDDDNSQDV or IE(pS)QEN(pS)KL(pS)QE, which has extremely high biosafety, calcium-chelating ability and calcium permeability.

[0012] The first object of the present invention is to provide a calcium-chelating peptide, and the amino acid sequence of the calcium-chelating peptide is DDLDDDDDNSQDV or IESQENSKLSQE.

[0013] In one embodiment, the serine in the calcium-chelating peptide can also be phospho-modified serine.

[0014] In one embodiment, the calcium-chelating peptide is DDLDDDDDNSQDV or DDLDDDDDN(pS)QDV or IE(pS)QEN(pS)KL(pS)QE; wherein, (pS) is phospho-modified serine.

[0015] In one embodiment, the preparation method of the calcium-chelating peptide includes solid-phase synthesis method / enzymatic hydrolysis method / microbial expression method.

[0016] The second object of the present invention is to provide a food, a medicine, a health product or a nutritional product, which contains an effective dose of any of the above-mentioned calcium chelating peptides;

[0017] Optionally, the food, the medicine, the health product or the nutritional product may also contain any of the above-mentioned calcium chelating peptides; the derivatives of the calcium chelating peptides refer to the calcium chelating peptide derivatives obtained by hydroxylating, carbonylating, carboxylating, methylating, acetylating, phosphorylating, esterifying or glycosylating the amino acid side chain groups, the amino terminus or the carbonyl terminus of the calcium chelating peptides.

[0018] In one embodiment, the medicine further contains pharmaceutically acceptable pharmaceutical excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field.

[0019] In one embodiment, the excipients include one or more of the following: binders such as cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol; fillers such as starch, sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid esters and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, etc.; lubricants such as hydrogenated vegetable oil, talc powder and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, polyvinylpyrrolidone, cellulose acetate phthalate; additionally, other adjuvants such as flavoring agents and sweetening agents may be added to the composition;

[0020] Optionally, the dosage forms of the medicine include, but are not limited to, oral dosage forms, injection dosage forms, inhalation dosage forms;

[0021] Optionally, the oral dosage forms include, but are not limited to, tablets, capsules, granules, oral liquids, oral suspensions;

[0022] Optionally, the injection dosage forms include, but are not limited to, injection solutions, injection powder injections;

[0023] Optionally, the inhalation dosage forms include, but are not limited to, aerosols, powder aerosols.

[0024] The third object of the present invention is to provide the application of any of the above-mentioned calcium chelating peptides in the preparation of a food, a medicine, a health product or a nutritional product, and the food, the medicine, the health product or the nutritional product is used to promote calcium absorption.

[0025] In one embodiment, the calcium-chelating peptide can also be used in the preparation of products for antioxidant, anti-inflammatory, cartilage repair and tissue regeneration (such as promoting bone matrix mineralization, enhancing bone density, accelerating bone tissue growth, shortening the fracture healing cycle), regulating metabolic and immune functions (such as regulating enzyme activity, enhancing immunity), preventing dental caries, and improving digestive function.

[0026] In one embodiment, the food includes, but is not limited to, cereal products, vegetable products, fruit products, meat products, seafood, egg products, dairy products, soy products, beverages;

[0027] The food also includes special dietary foods;

[0028] The health products also contain acceptable excipients.

[0029] In one embodiment, the food, medicine, health product or nutritional product is used to promote calcium absorption.

[0030] In one embodiment, the medicine also contains pharmaceutically acceptable medicinal excipients; the medicinal excipients refer to conventional drug carriers in the pharmaceutical field.

[0031] In one embodiment, the excipients include one or more of the following: binders such as cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol; fillers such as starch, sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, crospovidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbates, sorbitan fatty acids and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, etc.; lubricants such as hydrogenated vegetable oil, talc and polyethylene glycol; coating materials such as acrylic resin, hypromellose, polyvinylpyrrolidone, cellulose acetate phthalate; additionally, other adjuvants such as flavoring agents and sweetening agents can be added to the composition;

[0032] Optionally, the dosage form of the drug includes, but is not limited to, oral dosage forms, injection dosage forms, inhalation dosage forms;

[0033] Optionally, the oral dosage forms include, but are not limited to, tablets, capsules, granules, oral liquids, oral suspensions;

[0034] Optionally, the injection dosage forms include, but are not limited to, injection solutions, injection powder injections;

[0035] Optionally, the inhalation dosage forms include, but are not limited to, aerosols, powder aerosols;

[0036] Optionally, the food includes, but is not limited to, cereal products, vegetable products, fruit products, meat products, seafood, egg products, dairy products, soy products, and beverages; the food also includes special dietary foods; the health products also contain acceptable excipients.

[0037] Advantages of the present invention

[0038] The main reason for the binding of the peptide chain to calcium ions in the present invention is that the amino acid sequence contains a large number of acidic amino acids, and the free carboxyl groups on them can form coordination bonds with calcium ions. At the same time, the free amino groups also participate in the reaction to form soluble complexes, preventing calcium ion precipitation and increasing calcium absorption in the small intestine and its accumulation in the body.

[0039] The calcium-chelating peptides DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV, and IE(pS)QEN(pS)KL(pS)QE provided by the present invention are safe and non-toxic, have better physicochemical activities compared with traditional calcium supplements, and their calcium-chelating abilities reach 638.78 μg / mg, 446.95 μg / mg, and 419.36 μg / mg respectively. Moreover, they can all promote calcium transport in the Caco-2 cell monolayer, improve calcium absorption and bioavailability in the human body, and are used as calcium supplementation preparations. Description of the drawings

[0040] Figure 1 HPLC identification result of calcium-chelating peptide DDLDDDDDNSQDV;

[0041] Figure 2 LC-MS identification result of calcium-chelating peptide DDLDDDDDNSQDV;

[0042] Figure 3 HPLC identification result of calcium-chelating peptide IE(pS)QEN(pS)KL(pS)QE;

[0043] Figure 4 LC-MS identification result of calcium-chelating peptide IE(pS)QEN(pS)KL(pS)QE;

[0044] Figure 5 Effect of calcium-chelating peptide DDLDDDDDNSQDV on the viability of Caco-2 cells;

[0045] Figure 6 Calcium-chelating ability of calcium-chelating peptides; where VK7 is VLPVPQK, DV13 is DDLDDDDDNSQDV, pDV13 is DDLDDDDDN(pS)QDV, and IE12 is IE(pS)QEN(pS)KL(pS)QE;

[0046] Figure 7Effect of calcium-chelating peptides on calcium transport in Caco-2 cell monolayers; among them, VK7 is VLPVPQK, DV13 is DDLDDDDDNSQDV, and pDV13DDLDDDDDN(pS)QDV. Detailed implementation manners

[0047] The following are descriptions of the preferred embodiments of the present invention. It should be understood that the embodiments are for better explaining the present invention and are not used to limit the present invention.

[0048] Raw materials used in the embodiments:

[0049] LPN was isolated from fresh cow's milk. The specific method refers to the reference (Ma Ping, Sun Jie, Liu Ning. Isolation, purification and identification of osteopontin in cow's milk [J]. Food and Fermentation Industries, 2008, (06): 135-139.);

[0050] Pepsin, trypsin and bile salts were purchased from Sigma-Aldrich Company in the United States, and potassium chloride, sodium chloride and sodium hydroxide were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0051] All calcium-chelating peptides used in the embodiments were synthesized by Shanghai Kopeptide Biotechnology Co., Ltd.

[0052] Testing methods:

[0053] 1. HPLC method

[0054] High performance liquid chromatography (HPLC) was used. A Kromasil 100-5-C18 (4.6×250mm×5μm) chromatographic column was used, with 0.1% trifluoroacetic acid aqueous solution (A) and 0.1% trifluoroacetic acid acetonitrile solution (B) as the mobile phase. The total flow rate was 1 mL / min, and the detection wavelength was set at 214 nm. The sample was dissolved in ultrapure water, and the injection volume was 7 μL. The gradient elution program was as follows:

[0055] Time (min) Phase A (%) Phase B (%) 0.01 90 10 30 60 40 33 0 100 38 0 100 40 0 10 50 End

[0056] 2. LC-MS method

[0057] An electrospray ionization (ESI) interface was used. The detector voltage was set at -0.2 kV, the CDL temperature was 250 °C, the CDL voltage was 0 V, the block temperature was 200 °C, the atomizing gas flow rate was 1.5 L / min, the preset bias was +4.5 kV, and the T-Flow was 0.2 ml / min. The sample dissolution method was a mixed solvent of 15% acetonitrile (ACN) and 85% water (H2O), and the injection volume was 0.2 μL.

[0058] Example 1: Preparation of calcium-chelating peptides

[0059] A method for preparing calcium-chelating peptides, which uses an infant model for in vitro simulated digestion, includes the steps:

[0060] (1) Gastric digestion stage:

[0061] Add 10 mg / mL LPN solution to an enzyme reactor for preheating, adjust the pH to 5.3, and then mix it with simulated gastric juice (pH 5.3, 724.3 U pepsin / mL, 13 mM potassium chloride, 94 mM sodium chloride) at a ratio of 63:37 (v / v); react at 37 °C and pH 5.3 for 60 min, and adjust the pH to 7.0 with 2 M sodium hydroxide to terminate the digestion in this stage, obtaining the product of the gastric digestion stage;

[0062] (2) Intestinal digestion stage:

[0063] Adjust the pH of the product of the gastric digestion stage to 6.6, and then mix it with simulated intestinal juice (pH 6.6, 42.1 U trypsin / mL, 8.2 mM bile salts, 10 mM potassium chloride, 249 mM sodium chloride) at a ratio of 62:38 (v / v); digest at 37 °C and pH 6.6 for 60 min, and terminate the intestinal digestion at 95 °C for 10 min to obtain the final digestion product.

[0064] (3) Identification of polypeptide composition:

[0065] Analyze the final digestion product by LC-MS / MS equipped with an online nano-spray ion source. The whole system is an Orbitrap Fusion Lumos mass spectrometer (Thermo Fisher Scientific, MA, USA) in tandem with EASY-nanoLC 1200. A total of 5 μL of the sample is loaded (analysis column: Acclaim PepMap C18, 75 μm x 25 cm), the sample is separated with a gradient of 60 min, the column flow rate is controlled at 300 nL / min, the column temperature is 40 °C, the electrospray voltage is 2 kV, the gradient starts from 4% of phase B, rises to 50% in a non-linear gradient within 53 minutes and 40 seconds, rises to 95% in 40 seconds, and is maintained for 5 minutes and 40 seconds.

[0066] The mass spectrometer was operated in data-dependent acquisition mode, automatically switching between MS and MS / MS acquisitions. The mass spectrometry parameters were set as follows: (1) MS: scan range (m / z): 100 - 1500; resolution: 120,000; Normalized AGC target: 200%; maximum injection time: 100 ms; (2) HCD-MS / MS: resolution: 50,000; Normalized AGC target: 200%; maximum injection time: 86 ms; collision energy: 25%, 30%, 35%; dynamic exclusion time: 30 s. The tandem mass spectra were analyzed using PEAKS Studio version 10.6 (Bioinformatics Solutions Inc., Waterloo, Canada). The database was Uniprot-Bostaurus (version 2024, 26635 entries). The database search parameters were as follows: fragment ion mass tolerance: 0.02 Da, parent ion mass tolerance: 10 ppm, maximum missed cleavage: 2, fixed modification: Carbamidomethylation 57.02, variable modifications: Oxidation (M) 15.99, Deamidation (NQ) 0.98, Acetylation (Protein N-term) 42.01, Phosphorylation (STY) 79.97. The protein false discovery rate (FDR) cutoff was 1%, with at least 1 unique peptide; the peptide FDR cutoff was 1%.

[0067] A total of 242 peptides derived from LPN were identified, and three potential calcium-chelating peptides, DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV, and IE(pS)QEN(pS)KL(pS)QE, were selected based on the proportion of acidic amino acids and phosphorylation degree.

[0068] Shanghai Kopeptide Biotech Co., Ltd. was commissioned to synthesize the calcium-chelating peptides DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV, and IE(pS)QEN(pS)KL(pS)QE.

[0069] Example 2: Detection of calcium-chelating peptides

[0070] The calcium-chelating peptides DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV, and IE(pS)QEN(pS)KL(pS)QE prepared in Example 1 were taken, and the detection results were as follows:

[0071] (1) HPLC detection results

[0072] The HPLC detection results were as Figure 1 andFigure 3 as shown

[0073] (2) LC-MS detection results

[0074] The LC-MS detection results are as Figure 2 and Figure 4 shown

[0075] (3) Safety detection

[0076] To detect the biosafety of DDLDDDDDNSQDV and IE(pS)QEN(pS)KL(pS)QE, the steps are as follows:

[0077] Take Caco-2 cells in the logarithmic growth phase and inoculate 1.0×10 5 cells per well into a 96-well plate, 100 μL per well, and culture at 37 °C, 5% CO2 and saturated humidity for 24 h. Aspirate the culture medium, and add complete medium containing calcium chelating peptides at final concentrations of 0.1, 1, and 10 μg / mL to continue culturing the cells for 24 h. Use the cck-8 reagent to measure the cell viability.

[0078] The results are as Figure 5 shown. The results show that DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV and IE(pS)QEN(pS)KL(pS)QE at 0.1 - 10 μg / mL will not affect cell activity, that is, DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV and IE(pS)QEN(pS)KL(pS)QE have high biosafety.

[0079] (4) Calcium chelating ability

[0080] To detect the calcium chelating ability of DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV and IE(pS)QEN(pS)KL(pS)QE, the steps are as follows:

[0081] Mix the freeze-dried powder (2 mg) of DDLDDDDDNSQDV, DDLDDDDDN(pS)QDV and IE(pS)QEN(pS)KL(pS)QE with 1 ml of 5 mM CaCl2 solution and 2 mL of phosphate buffer (pH 7.8, 20 mM), shake well until completely dissolved. Stir at 37 °C for 120 min, centrifuge the reaction solution at 4000 g for 20 min to precipitate insoluble calcium phosphate salts.

[0082] The calcium content in the supernatant was determined by flame atomic absorption spectrometry. Deionized water was used instead of the sample as a blank control, and calcium = soluble calcium in the supernatant of the peptide-containing sample - soluble calcium in the blank supernatant. The experiment was repeated three times for each group, and the average value ± standard deviation was used to represent the results. The polypeptide VLPVPQK disclosed in the existing literature was used as a control.

[0083] The results are as Figure 6 shown. The results indicate that the calcium chelating ability of DDLDDDDDNSQDV can reach 638.78 μg / mg, that of DDLDDDDDN(pS)QDV can reach 446.95 μg / mg, and that of IE(pS)QEN(pS)KL(pS)QE can reach 419.36 μg / mg, all of which are significantly higher than that of VK7 (342.81 μg / mg).

[0084] (5) Effects on calcium transport

[0085] To detect the effects of calcium-chelating peptides on calcium transport in a monolayer of Caco-2 cells, the steps are as follows:

[0086] Adjust the concentration of the Caco-2 cell suspension to 5×10 4 cells / mL and inoculate it on the filter membrane of a 12-well Transwell plate. Add 0.5 mL of the cell suspension and 1.5 mL of complete DMEM culture medium to the apical side (AP) and basolateral side (BL) of the filter membrane, respectively. After 24 h, change the culture medium. Change the medium every other day for the first 7 days and then every day until the cells form a tight monolayer.

[0087] Take the Transwell plate inoculated with Caco-2 cells out of the cell culture incubator, equilibrate it at room temperature in a laminar flow hood for 30 min, and then measure the transepithelial electrical resistance (TEER) of the cell membrane using a Millipore Millicell-ERS epithelial transmembrane cell resistance meter. The test electrodes need to be sterilized by soaking in 70% alcohol for 15 min, taken out and air-dried for 15 s, then rinsed with sterile PBS and then with complete culture medium. When measuring the transmembrane resistance, immerse the short end of the electrode in the AP side and the long end in the BL side. When TEER > 500 Ω·cm 2 it can be used for intestinal transport experiments.

[0088] After the formation of the Caco-2 cell monolayer, the Caco-2 cells in the Transwell plate were washed twice with HBSS buffer. When adding HBSS for the third time, the culture plate was placed in an incubator at 37 °C and incubated for 30 min. Then, the HBSS in the wells was gently aspirated. On the AP side, 0.5 mL of the working solution (containing 10 μg / mL calcium chelating peptide and 5 mM CaCl2 prepared with HBSS) was added, and 0.5 mL of HBSS containing 5 mM CaCl2 was added to the control group. On the BL side, 1.5 mL of HBSS buffer was added. After 2 h of transport, the samples on the BL side were taken out. The calcium content of the samples was determined by flame atomic absorption spectrometry. Calcium permeability (%) = (calcium content on the BL side / initial calcium content on the AP side) * 100%.

[0089] The results are as Figure 7 shown. The results indicate that the total calcium permeability of the DDLDDDDDNSQDV group was 36.53%, that of the DDLDDDDDN(pS)QDV group was 39.12%, and that of the IE(pS)QEN(pS)KL(pS)QE group was 40.79%, all of which were significantly higher than 11.20% of the control group and 34.55% of the VK7 group.

[0090] Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any person familiar with this technology can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.

Claims

1. A calcium-chelating peptide, characterized in that, The amino acid sequence of the calcium chelating peptide is DDLDDDDDNSQDV or IESQENSKLSQE.

2. The calcium-chelating peptide according to claim 1, characterized in that, The serine in the calcium chelating peptide may also be phosphorylated serine.

3. The calcium-chelating peptide according to any one of claims 1 to 2, characterized in that, The preparation method of the calcium chelating peptide includes solid-phase synthesis method / enzyme digestion method / microbial expression method.

4. A food, medicine, health product or nutritional product, characterized in that, The food, drug, health product or nutritional product contains an effective dose of the calcium chelating peptide according to any one of claims 1 to 3; Optionally, the food, drug, health product or nutritional product may also contain the calcium chelating peptide according to any one of claims 1 to 3; the derivative of the calcium chelating peptide refers to the calcium chelating peptide derivative obtained by hydroxylating, carbonylating, carboxylating, methylating, acetylating, phosphorylating, esterifying or glycosylating the amino acid side chain group, amino terminus or carbonyl terminus of the calcium chelating peptide.

5. The food, drug, health product or nutritional product according to claim 4, characterized in that, The drug also contains a pharmaceutically acceptable pharmaceutical excipient; the pharmaceutical excipient refers to a conventional drug carrier in the pharmaceutical field.

6. The food, medicine, health product or nutritional product according to claim 5, characterized in that, The excipient includes one or more of the following: binders such as cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol; fillers such as starch, sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid esters and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, etc.; lubricants such as hydrogenated vegetable oil, talc powder and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, polyvinylpyrrolidone, cellulose acetate phthalate; additionally, other adjuvants such as flavoring agents and sweetening agents may be added to the composition; Optionally, the dosage form of the drug includes but is not limited to oral dosage form, injection dosage form, inhalation dosage form; Optionally, the oral dosage form includes but is not limited to tablets, capsules, granules, oral liquids, oral suspensions; Optionally, the injection dosage form includes but is not limited to injection solutions, injection powder for injection; Optionally, the inhalation dosage form includes but is not limited to aerosols, powder aerosols.

7. The food, drug, health product or nutritional product according to claim 4, characterized in that, The food includes but is not limited to cereal products, vegetable products, fruit products, meat products, seafood, egg products, dairy products, soy products, beverages; The food also includes special dietary foods; The health product also contains an acceptable excipient.

8. Use of the calcium chelating peptide according to any one of claims 1 to 3 in the preparation of food, medicine, health food or nutritional product, characterized in that, The food, drug, health product or nutritional product is used to promote calcium absorption.

9. The application according to claim 8, wherein The drug also contains a pharmaceutically acceptable pharmaceutical excipient; the pharmaceutical excipient refers to a conventional drug carrier in the pharmaceutical field.

10. The application according to claim 9, wherein The excipients include one or more of the following: binders such as cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol; fillers such as starch, sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, and dried starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid esters, and glycerol fatty acid esters; coloring agents such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, etc.; lubricants such as hydrogenated vegetable oil, talc, and polyethylene glycol; coating materials such as acrylic resin, hypromellose, polyvinylpyrrolidone, cellulose acetate phthalate; additionally, other adjuvants such as flavoring agents and sweetening agents can be added to the composition. Optionally, the dosage form of the drug includes, but is not limited to, oral dosage forms, injection dosage forms, inhalation dosage forms. Optionally, the oral dosage forms include, but are not limited to, tablets, capsules, granules, oral liquids, oral suspensions. Optionally, the injection dosage forms include, but are not limited to, injection solutions, injection powder for injection. Optionally, the inhalation dosage forms include, but are not limited to, aerosols, powder inhalants. Optionally, the food includes, but is not limited to, cereal products, vegetable products, fruit products, meat products, seafood, egg products, dairy products, soy products, beverages; the food also includes special dietary foods; the health products also contain acceptable excipients.

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