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Preparation method and application of sinonovacula constricta enzymolysis polypeptide

A technology of enzymatic hydrolysis and razor clam, which is applied in the field of preparation of enzymatic hydrolyzed polypeptides of razor clam, achieving the effects of high sensitivity, good stability, and few side effects

Inactive Publication Date: 2014-05-21
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Preparation method and application of sinonovacula constricta enzymolysis polypeptide

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Embodiment Construction

[0031] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0032] 1. Materials and Instruments

[0033] 1.1 Main materials and reagents

[0034] Hanging razor clams were purchased from Nanzhen Market, Zhoushan City, Zhejiang Province;

[0035] Fetal bovine serum (FBS), Hangzhou Sijiqing Biological Products Co., Ltd.;

[0036] HamF12 medium, Gibco company;

[0037] Trypsin, American SIGMA company;

[0038] Penicillin, streptomycin, Shandong Lukang Pharmaceutical Co., Ltd.;

[0039] Dextran gel Sephadex G-25, Beijing Asia Pacific Hengxin Biotechnology Co., Ltd.;

[0040] Pepsin, papain, alkaline protease, Asia-Pacific Hengxin Biotechnology Co., Ltd.;

[0041] MTT was purchased from Sigma, USA;

[0042] Dimethyl sulfoxide (DMSO) was purchased from American AMRESCO;

[0043] The rest of the reagents were analytically pure.

[0044] 1.2 Main instruments

[0045] DS-1 high-speed tissue grinder, Shan...

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Abstract

The invention discloses a preparation method and application of a sinonovacula constricta enzymolysis polypeptide. The method is characterized by comprising the following steps: shelling sinonovacula constricta, washing, mincing, adding water in the material-liquid mass ratio of 1:(3-5) for homogenizing, and adjusting the pH value to 5.5-6.5; adding protease for performing enzymolysis at the temperature of 60-70 DEG C for 1.5-2.5 hours, wherein the adding amount of the protease is 0.2-0.3 percent based on the total mass of sinonovacula constricta; after enzymolysis, performing enzyme deactivation, and centrifuging to obtain supernatant serving as enzymatic hydrolysate; performing ultrafiltration, G-25 gel chromatography and reversed phase high-performance liquid chromatography elution on the enzymatic hydrolysate to obtain the needed sinonovacula constricta enzymolysis polypeptide. A preparation process is simple, the obtained sinonovacula constricta enzymolysis polypeptide is high in sensitivity and stability and small in side effects, plays a remarkable role in inhibiting the proliferation of DU-145 and PC-3 cells through MTT detection, and can be applied to in-vitro resistance of prostatic cancer. The preparation method plays an important role in further researching and developing functional foods and medicaments based on the sinonovacula constricta enzymolysis polypeptide and increasing the economical value of the sinonovacula constricta.

Description

technical field [0001] The invention relates to a preparation method of a razor clam enzymolysis polypeptide and its application in anti-prostate cancer. 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 Petalbranchia, Heterodontia, Pleurophyllidae, Lighthouse Clamidae, and genus Constricted razor clam. It is distributed in the coastal areas of my country and has a large output. It is one of the four major cultured shellfish in my country[ An Xianhui, Li Liantai. Research status and development prospect of razor clam [J]. Scientific Fish Culture, 2005 (1): 4-6.] The meat of razor clam is delicious and nutritious, rich in various essential amino acids and unsaturated fatty acids required by human life activities Nutrients [Li Taiwu, Lin Ye, Su Xiurong, Multivariate...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/20C07K1/16A61K38/01A61P35/00
Inventor 丁国芳李丽黄芳芳闫海强
Owner ZHEJIANG OCEAN UNIV
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