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Method for preparing calcium chelated peptide

A calcium chelation and chelation technology, applied in the field of functional peptide preparation, can solve the problems of low yield of target peptide, high cost, cumbersome operation steps, etc.

Active Publication Date: 2014-12-24
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Scholars at home and abroad have isolated polypeptides with metal ion chelating activity from different sources of active protein hydrolyzate, mainly collagen oligopeptides, casein phosphopeptides, phosvitin phosphopeptides, etc., but most metal ions Chelating peptides are all prepared by chromatographic separation and affinity chromatography. The operation steps are cumbersome, the yield of the target peptide is low, and the cost is high. There is an urgent need for a new technology for the preparation of metal ion chelating peptides.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Cod skin collagen preparation method is as follows: Soak cod skin with 0.05mol / L hydrochloric acid solution (solid-liquid ratio: 1 / 6) for 40 minutes, wash until neutral, leaching with hot water at 75°C (solid-liquid ratio: 1 / 2) for 4 hours, The fish skin is removed by filtration, the obtained solution is centrifuged to remove impurities, and the supernatant is freeze-dried to obtain the cod skin collagen.

[0022] Take the collagen extracted from cod skin, add water until the protein concentration is 2%, add sodium tripolyphosphate to 1 / 3 of the collagen, adjust the pH to 8, and the reaction temperature is 80°C. Auxiliary treatment therefore prolongs the reaction time, and the reaction takes 2.5 hours. After the reaction, pour cooling water into the interlayer of the reaction kettle, cool down to below 8°C, and pump it into the precipitation tank; pump saturated ammonium sulfate solution into the precipitation tank under stirring until Stop when the concentration of amm...

Embodiment 2

[0027] Take collagen, add water until the protein concentration is 3%, add sodium tripolyphosphate to 1 / 3 of the collagen, adjust the pH to 8, and the reaction temperature to 80°C, use high-voltage pulse electric field treatment for auxiliary treatment, 500 pulses , pulse time 150μs, frequency 100Hz, react for 1.5h, pass cooling water into the interlayer of the reactor, cool down to below 8°C, pump into the precipitation tank; pump saturated ammonium sulfate solution into the precipitation tank under stirring until the salting out solution Stop when the concentration of ammonium sulfate in the medium reaches 35%, centrifuge after standing for 30min, centrifuge at 10000r / min for 10min, discard the supernatant, and wash the sediment several times with ammonium sulfate solution with a saturation of 75%, centrifuge to take the precipitate, and The precipitate was dissolved and ultrafiltered with a 2000 Dalton hollow fiber ultrafiltration membrane to obtain desalted phosphorylated c...

Embodiment 3

[0032]Weigh 1kg of collagen and place it in a reaction kettle, dilute it to a protein concentration of 3% with distilled water, add sodium tripolyphosphate to 1 / 3 of the collagen, adjust the pH to 8.5, and the reaction temperature to 80°C. The high-voltage pulse electric field treatment is used as an auxiliary treatment, so the reaction time is prolonged, and the reaction takes 3 hours. After the reaction, the cooling water is passed into the interlayer of the reaction kettle, the temperature is lowered to below 8°C, and the pump is pumped into the precipitation tank; the saturated sulfuric acid is pumped into the precipitation tank under stirring. Ammonium solution, until the concentration of ammonium sulfate in the salting-out solution reaches 35%, let it stand for 50 minutes, then centrifuge in a freezer, centrifuge at 10000r / min for 10 minutes, discard the supernatant, and wash the precipitate fully with ammonium sulfate solution with a saturation of 75% again Several times...

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Abstract

The invention aims at providing a method for preparing calcium chelated peptide. The method comprises the following steps: firstly, phosphorylating a collagen solution at 70-85 DEG C by using a phosphorylation agent, purifying and desalting the phosphorylated collagen, and adding Alcalase into the prepared desalted and phosphorylated collage solution to perform enzymolysis with a trypsin complex enzyme line so as to prepare the calcium chelated peptide. The hydrolysis degree of phosphorylated collagen peptide prepared by using the method is 14-20%, the peptide chain length range of the phosphorylated collagen peptide is within 5-10 amino groups, the phosphorus content of the phosphorylated collagen peptide is 91-138 mu g of phosphorus in every gram of collagen, and the phosphorylated collagen peptide is good in chelation property with calcium ions. Compared with conventional collagen, the calcium chelated peptide is better in physiological activity, the absorption and the utilization of human bodies on calcium ions are promoted, and a phosphorylation process is applicable to industrialization production.

Description

technical field [0001] The invention belongs to the technical field of preparation of functional polypeptides, and in particular relates to a preparation method of phosphorylated collagen peptides capable of chelating with calcium ions. Background technique [0002] Due to the relatively high proportion of plant foods in my country's dietary structure, the intake of dairy products is relatively small, and calcium deficiency often occurs, especially in some children and the elderly. Studies have shown that calcium deficiency is caused by insufficient calcium intake on the one hand, and insufficient absorption of calcium in the body on the other hand. Many studies have confirmed that the absorption of calcium in the small intestine is affected by many factors, such as the amount of dietary calcium intake and its presence, the influence of other ingredients in the diet, the absorption function of the body itself, and environmental factors. The body's apparent absorption rate o...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/78C07K1/30
Inventor 侯虎李八方彭喆张朝辉赵雪陈铁军
Owner OCEAN UNIV OF CHINA