Preparation method of sea cucumber ovum peptide-calcium chelate

A sea cucumber egg and chelate technology, which is applied in the field of preparation of sea cucumber egg peptide calcium chelate, can solve the problem that the calcium absorption rate is less than 40%, and achieve the effects of simple operation process, reduced pollution, and favorable calcium absorption

Active Publication Date: 2016-06-15
DALIAN POLYTECHNIC UNIVERSITY
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Problems solved by technology

[0004] The inorganic calcium and organic calcium preparations currently on the market will partially form precipitates in the weak alkaline environment of the small intestine, resulting in a calcium absorption rate of less than 40%.
There is no relevant literature report on the preparation of peptide calcium chelate by using biological enzyme method to obtain bioactive peptides from sea cucumber eggs

Method used

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  • Preparation method of sea cucumber ovum peptide-calcium chelate
  • Preparation method of sea cucumber ovum peptide-calcium chelate
  • Preparation method of sea cucumber ovum peptide-calcium chelate

Examples

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Effect test

Embodiment 1

[0025] 1. Sea cucumber eggs were thawed in running water, rinsed with deionized water, homogenized, boiled in a boiling water bath for 5 minutes for denaturation pretreatment, and sea cucumber eggs homogenate was obtained;

[0026] 2. Take the sea cucumber egg homogenate, add 10 times the volume of n-hexane / absolute ethanol (3:1) mixed solution 100mL, extract at 50°C for 6 hours, repeat 1-2 times, remove fat, and the filter cake obtained after suction filtration is natural Air-dried to obtain degreased sea cucumber egg powder, and adopt the Kjeldahl nitrogen analyzer to measure the protein content;

[0027] 3. Accurately weigh the degreased sea cucumber egg powder, add water until the substrate protein concentration is 2g / 100mL, add trypsin, alkaline protease, neutral protease, papain, flavor protease respectively under the optimum enzymatic hydrolysis conditions (in this patent The optimum condition of the enzyme is determined according to the optimum condition of the enzyme ...

Embodiment 2

[0032] Embodiment 2: the mensuration of peptide yield

[0033] 1. Prepare 0.5mg / mL bovine serum albumin (BSA) standard solution: Accurately weigh 0.05gBSA in a 100mL clean beaker with a precision analytical balance, dissolve it in ultrapure water, and set the volume to 100mL.

[0034] 2. Draw the BSA standard curve by Folin-phenol method: In this experiment, BSA was used as the standard to draw the standard curve. Take 18 2.0mL centrifuge tubes and divide them into 3 groups. Add 0, 20, 40, 60, 80, and 100 μL standard solution respectively, and make up to 100 μL with ultrapure water, and do 3 parallels for each concentration above. The concentration of each tube is 0, 0.1, 0.2, 0.3, 0.4, 0.5 mg / mL in turn. Add 500 μL of Folin-phenol A solution to each tube sequentially and mix well. Place at room temperature for 10 minutes, then add 50 μL of Folin-phenol B solution, shake evenly immediately, and place at room temperature for 30 minutes. Take 200 μL of the solution from each...

Embodiment 3

[0040] Embodiment 3: the preparation of sea cucumber egg peptide calcium chelate

[0041] 1. Combine enzymatic hydrolyzate of sea cucumber eggs with CaCl 2Solution and phosphate buffer were mixed to prepare a chelation reaction solution, in which the final concentration of the enzymatic hydrolyzate was 0.5mg / mL, CaCl 2 The final concentration was 5mmol / L, and the final concentration of phosphate buffer was 20mmol / L; the prepared chelation reaction solution was subjected to chelation reaction under simulated ileal conditions (37°C, pH7.0) for 1h, and the pH was maintained at 7.0 ( pH meter monitoring, 1mol / LHCl or NaOH adjustment);

[0042] 2. After the chelation reaction, the mixed solution was centrifuged at 5000×g for 10 minutes to remove the calcium phosphate precipitate, and the supernatant was passed through a 0.45 μm aqueous microporous filter membrane, and freeze-dried to obtain the sea cucumber egg peptide calcium chelate.

[0043] Calcium binding was determined usin...

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Abstract

The invention discloses a preparation method of sea cucumber ovum peptide-calcium chelate. The preparation method comprises the following steps: pretreating sea cucumber ova; skimming; performing enzymolysis; simulating an ileum condition to obtain the sea cucumber ovum peptide-calcium chelate. By adopting the method, the utilization ratio of byproducts produced in a sea cucumber processing process is increased, contained active substances are developed and utilized fully, and meanwhile environmental pollution is lowered; sea cucumber ovum biological active peptide is prepared by a biological enzyme method, and soluble peptide calcium chelate formed by means of combination with calcium ions contributes to calcium absorption and can be used for developing a high-valued sea cucumber ovum peptide-calcium supplement; an operation process is simple, and complex equipment is not needed; the peptide yield and calcium combining quantity of sea cucumber ovum peptide obtained by trypsin enzymolysis are highest, the peptide yield is up to 86 percent, and the calcium combining quantity is 0.38mmol / L; moreover, high calcium absorption prompting capability is achieved, and the production efficiency is increased; the preparation method is suitable for industrial production.

Description

technical field [0001] The invention relates to the field of preparation of sea cucumber-based functional active substances, and more specifically relates to a preparation method of sea cucumber egg peptide calcium chelate. Background technique [0002] In recent years, China's sea cucumber industry has developed rapidly. According to the 2015 Fisheries Yearbook, China's total sea cucumber production is about 200,000 tons. Sea cucumber eggs are one of the by-products of sea cucumber processing. However, the recycling of sea cucumber eggs has been ignored by people. They are usually regarded as industrial waste, which is not only a waste of biological resources, but also causes environmental pollution. [0003] Calcium is an essential constant element for the human body. It is not only an important component of bones and teeth, but also a regulator of metabolic activities in the body. It participates in physiological activities such as muscle contraction, nerve conduction, bl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
CPCC12P21/06
Inventor 孙娜崔蓬勃吴海涛于翠平唐越阎佳楠查越
Owner DALIAN POLYTECHNIC UNIVERSITY
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