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A kind of clam clam active octapeptide and its preparation method and application
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A constricted and active technology, which is applied in the field of constricted active octapeptide and its preparation, and achieves the effects of mild enzymatic hydrolysis, wide source of raw materials, and inhibiting cell proliferation activity.
Active Publication Date: 2020-11-06
ZHEJIANG OCEAN UNIV
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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.
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Embodiment 1
[0031] Embodiment 1: Preparation of active octapeptide of razor clam constriction
[0032] 1. Pretreatment:
[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 Alkal...
Embodiment 2
[0063] Example 2: Study on the anti-proliferation activity of razor clam active octapeptide on 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 octapeptide 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 Compa...
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Abstract
The invention relates to active octapeptide of sinonovacula constricta. The amino acid sequence of the active octapeptide of sinonovacula constricta is as follows: Met-His-Thr-Asp-Asp-Asp-Val-Glue, and the molecular weight detected by ESI / MS is 960.10Da. A preparation method comprises the following steps: unshelling fresh sinonovacula constricta and fetching meat, triturating cleaned sinonovacula constricta meat for tissue homogenation, and adjusting the pH value of a homogenization liquid after homogenization for later use; and adding pepsase into the homogenization liquid for enzymolysis, carrying out ultrafiltration and reversed phase high performance liquid chromatography purification of G-25 gel separation on an obtained enzymolysis sample, and collecting an eluting peak component for amino acid sequencing to obtain the active octapeptide of sinonovacula constricta. The maximum proliferation inhibiting rate of the active octapeptide of sinonovacula constricta on a human prostatic cancer PC-3 cell can reach over 90%, can provide a theoretical basis for researching anti-prostate cancer drugs, and is of significance in further researching and developing drugs and foods based on the active octapeptide of sinonovacula constricta and improving the economical value of the active octapeptide of sinonovacula constricta.
Description
technical field [0001] The invention relates to the field of bioactive peptides, in particular to an active octapeptide 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 Petalbranchia, Heterodontia, Pleurophyllidae, Lighthouse Clamidae, and 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. 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 Taiwu, Lin Y...
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