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Preparation method and application of Cyclina sinensis enzymolysis polypeptide

A green clams enzyme and a technology for clams, which are applied in the field of preparation of clams enzymolyzed polypeptides, can solve problems such as no reports on clams enzymolyzed polypeptides, and achieve the effects of low cost, convenient operation and wide sources.

Inactive Publication Date: 2015-07-22
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current studies have shown that the extract of clam meat can increase the activity of lymphocytes and macrophages in the spleen of aged rats and the activity of macrophages in lymph nodes, which plays an important role in improving cellular immune response and normal immunity of the body; crude Clam polysaccharides have antioxidant activity in vitro, and have an important protective effect on the carbon tetrachloride-induced hepatotoxicity model in mice, but there is no report on the enzymatic polypeptide of Clam, especially in the role in antioxidant activity

Method used

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  • Preparation method and application of Cyclina sinensis enzymolysis polypeptide
  • Preparation method and application of Cyclina sinensis enzymolysis polypeptide
  • Preparation method and application of Cyclina sinensis enzymolysis polypeptide

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1: Enzymolysis preparation of green clam enzymolysis polypeptide

[0031] The viscera of the above-mentioned green clam is taken as a raw material, and the raw material is crushed with a homogenizer to obtain a homogenate liquid of the viscera of the green clam. Add distilled water to the above homogenate at a ratio of 1:3 to 4, adjust the pH value of the homogenate with a certain concentration of hydrochloric acid and sodium hydroxide solution, add alkaline protease for enzymolysis, and inactivate it with boiling water for 10~ 15min. Cool the inactivated enzymatic solution to room temperature, centrifuge at 9,000-10,000 r / min for 10-15 minutes, take the supernatant, concentrate it by rotary evaporation, and freeze-dry to obtain the clam enzymatic polypeptide.

[0032] As shown in Table 1, considering the four factors in the enzymolysis process: 1. temperature, 2. time, 3. the amount of enzyme added, and 4. pH, the influence on the effect of enzymolysis, each f...

Embodiment 2

[0038] Example 2: The scavenging rate of DPPH free radicals by enzymatic hydrolyzed polypeptide of green clam

[0039] The hydrolyzate obtained under the best enzymolysis conditions obtained in Example 1 was freeze-dried, and dissolved in distilled water to obtain a test sample. The mass concentration of the test sample was 1mg / mL, 2mg / mL, 3mg / mL, 4mg / mL , 5mg / mL. The DPPH concentration is 0.1mmol / L (dissolved in absolute ethanol) and stored in the dark. The method is 1mL sample + 1mL DPPH. After mixing, store in the dark for 30min, centrifuge at 4000r / min for 10min, and measure the absorbance Ai at 517nm. Simultaneously measure the absorbance Aj of 1mL absolute ethanol + 1mL sample mixture, and measure the absorbance Ac by mixing with 1mL DPPH + 1mL water. The calculation formula is: DPPH clearance rate (%)=[1-(Ai-Aj) / Ac]×100 %.

[0040] The result is as figure 1 As shown, it can be seen that the scavenging rate of the clam alkaline protease hydrolyzate to DPPH free radica...

Embodiment 3

[0041] Example 3: Hydroxyl free radical scavenging rate of clam enzymatic hydrolyzed polypeptide

[0042] Take 1mL of 0.75mmol / L phenanthroline in a test tube, add 2mL of PBS with a pH value of 7.4, 1mL of sample solution in turn, shake and mix well, add 1mL of ferrous sulfate with a concentration of 0.75mol / L, mix well, and then Add 1 mL of 0.03% hydrogen peroxide, bathe in 37°C water for 1 h, measure its absorbance As at 536nm; the blank group replaces hydrogen peroxide with water, which is Ab; (Ab-Ap)] × 100%, sample processing is the same as in Example 2.

[0043] The result is as figure 2As shown, it can be seen that alkaline protease hydrolyzates at different concentrations have a certain degree of scavenging effect on hydroxyl free radicals, and as the concentration increases, the scavenging rate increases, but the scavenging rate of enzymatic polypeptides on hydroxyl free radicals is lower than that of DPPH , when the concentration is 5mg / mL, the scavenging rate is ...

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Abstract

The invention relates to a preparation method of Cyclina sinensis enzymolysis polypeptide. The method includes: subjecting fresh Cyclina sinensis to evisceration and homogenization, taking the homogenate, adding alkaline protease to perform enzymolysis, centrifuging the enzymolysis solution and taking the supernatant, carrying out rotary evaporation and concentration, and freeze drying so as to obtain Cyclina sinensis enzymolysis polypeptide with a molecular weight of 15-25kDa. The Cyclina sinensis enzymolysis polypeptide has good antioxidant activity, has a DPPH free radical clearance rate up to 91.24%, a hydroxyl free radical clearance rate up to 62.83%, chelating power to Fe<2+> of 61.13%, and reducing power of 0.331, and also has certain protective effect on hydroxyl free radical induced damaged pBR322 DNA, and can be widely used in antioxidation active products.

Description

technical field [0001] The invention relates to the technical field of marine organisms, in particular to a preparation method and application of a clam enzymatically hydrolyzed polypeptide. Background technique [0002] Green clams (Cyclina sinensis) belong to the mollusk phylum, petal branch class, clam order, and clam family, commonly known as black clam, round clam, iron clam, etc. A common shellfish. Green clams are delicious and nutritious. Besides their high edible value, they are also an important marine medicine. "Shen Nong's Materia Medica", "Compendium of Materia Medica" and "Supplements to Compendium of Materia Medica" all use clams as a commonly used medicine. included in it. [0003] In recent years, there have been many studies on enzymatic polypeptides of seafood, such as: the Chinese invention patent "Preparation Method of Enzymatic Polypeptides of Philippine Clams" with patent number ZL201010253745.0 (authorized announcement number CN 102372765 B) disclos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
Inventor 杨最素丁国芳潘欣赵莎莎赵玉勤闫海强
Owner ZHEJIANG OCEAN UNIV
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