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Active decapeptide of Sinonovacula constricta and preparation method therefor and application thereof

A constricted and active technology, which is applied in the field of constricted active decapeptide and its preparation, and achieves the effects of mild enzymatic hydrolysis, inhibiting cell proliferation activity and wide sources.

Active Publication Date: 2015-11-25
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For razor clam constriction, there are few studies on anti-tumor aspects, especially the anti-tumor studies on the use of enzymatic hydrolysis technology to extract polypeptides have not been reported.

Method used

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  • Active decapeptide of Sinonovacula constricta and preparation method therefor and application thereof
  • Active decapeptide of Sinonovacula constricta and preparation method therefor and application thereof
  • Active decapeptide of Sinonovacula constricta and preparation method therefor and application thereof

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of active decapeptide of razor clam constriction

[0032] 1. Pre-processing:

[0033] Take fresh razor clam, remove the shell and take the meat of razor clam, wash it and drain it for later use.

[0034] 2. Pepsin hydrolysis

[0035] 2.1 Use a high-speed tissue masher to mash the razor clam meat, add distilled water to homogenate, the ratio of material to liquid is 1:1~3, accurately weigh the homogenate, and adjust it with 0.1mol / L hydrochloric acid solution and 0.1mol / L NaOH solution For the pH value of the homogenate, add pepsin for enzymolysis for several hours. The enzymolysis conditions are: enzymolysis temperature 30-45°C, pH 2-3.5, enzymolysis time 2-8 hours, enzyme amount 300-1200U / g razor clam meat. After the enzymatic hydrolysis, inactivate the enzyme at 100°C for 15 minutes, centrifuge at 4°C for 15 minutes (10000r / min), and take the supernatant for later use.

[0036] 2.2 Pepsin hydrolysis process optimization

[0037] 2.2.1...

Embodiment 2

[0063] Example 2: Research on the anti-proliferation activity of razor clam active decapeptide in PC-3 cells

[0064] Taking the F12 medium as the control group, the above-mentioned MTT method was used to compare the effects of different concentrations and different action times of active tripeptides on the proliferation of PC-3 cells. The test results are as follows: Figure 4 As shown, it can be seen that the active decapeptide of razor clam can effectively inhibit the proliferation of PC-3 in a concentration- and time-dependent manner.

[0065] Main materials and reagents used in the above-mentioned each embodiment:

[0066] Hanging razor clams were purchased from Nanzhen Market, Zhoushan City, Zhejiang Province; prostate cancer cell PC-3 hormone-independent cells (originally purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences and preserved by our laboratory); fetal bovine serum (FBS), Hangzhou Sijiqing Biological Products Ltd.; HamF12 medium, Gibco Co...

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Abstract

The invention relates to an active decapeptide of Sinonovacula constricta. According to the active decapeptide of Sinonovacula constricta, the amino acid sequence is Ile-Asp-Gly-His-Ala-Ala-Gly-Asp-Val-Gly, and shown by ESI / MS detection, the molecular weight is 835.7Da. The active decapeptide of Sinonovacula constricta is prepared through the following steps: shelling fresh Sinonovacula constricta to take flesh, washing the Sinonovacula constricta flesh, then, smashing the washed Sinonovacula constricta flesh, carrying out tissue homogenizing, and then, adjusting the pH value of the homogenate for later use; and adding pepsase into the homogenate for enzymolysis, carrying out ultrafiltration, G-25gel separation and out-phase high performance liquid chromatography purification on the obtained enzymolysis sample, collecting an elution peak component, and carrying out amino acid sequencing, thereby obtaining the active decapeptide of Sinonovacula constricta. According to the active decapeptide of Sinonovacula constricta, the maximum proliferation inhibition rate to human prostatic carcinoma PC-3 cells can reach over 90%, a theoretical basis can be provided for research on prostatic carcinoma resisting drugs, and the active decapeptide of Sinonovacula constricta has great significance in further researching developing drugs and food based on the active decapeptide of Sinonovacula constricta and increasing the economic value of the active decapeptide of Sinonovacula constricta.

Description

technical field [0001] The invention relates to the field of bioactive peptides, in particular to an active decapeptide of razor clam constriction and its preparation method and application. Background technique [0002] Marine shellfish is rich in resources and rich in many physiologically active substances, so it has been widely studied and applied gradually. Constricted razor clam (Sinonovacula constricta), commonly known as razor clam, belongs to the class Petalbranchia, Heterodonta, Pleurophyllidae, Lighthouse Clamidae, and the genus Constricted razor clam. It is distributed along the coast of my country and has a large output. It is one of the four major cultured shellfish in my country. Virtuous, Li Liantai. Research status and development prospect of razor clam. Scientific Fish Culture, 2005 (1): 4-6.], razor clam meat is delicious, nutritious, and rich in various essential amino acids, unsaturated fatty acids and other nutrients needed by human life activities[ Li ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C12P21/06C07K1/34C07K1/20A61K38/08A61P35/00A23L1/305
Inventor 黄芳芳丁国芳杨最素余方苗
Owner ZHEJIANG OCEAN UNIV
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