Preparation method of tuna extract with inhibitory effect on cancer cells

A technology of tuna and extract, which is applied in the field of preparation of tuna extract, can solve the problems of low enzymatic hydrolysis efficiency and achieve the effects of high enzymatic hydrolysis efficiency, reduced motility, and low preparation cost

Inactive Publication Date: 2017-10-20
浦江县泰如食品科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Marine biological proteins are very different from terrestrial biological proteins in terms of amino acid composition and amino acid sequence. The above method uses protease extracted from land to enzymatically hydrolyze marine organisms, and the enzymatic hydrolysis efficiency is low.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The preparation method of tuna extract with the effect of inhibiting cancer cells, including complex enzyme enzymolysis, isoelectric point precipitation, ultrafiltration and nanofiltration, HPLC elution and freeze-drying; Tuna meat homogenate was hydrolyzed with protease and wakame protease, and the hydrolysis was assisted by ultrasonic waves. Marine biological proteins are very different from terrestrial biological proteins in terms of amino acid composition and amino acid sequence. Marine Brevibacterium oxidans produces alkaline protease and wakame protease to enzymatically hydrolyze tuna protein at the same time, and a large number of active peptides can be obtained .

[0020] The tuna extract contains an active short peptide with the amino acid sequence HSHACASYYCRCLVLHPCVNCYCRCSR. The above-mentioned active short peptide can effectively inhibit the unwinding of the double helix structure of cancer cell DNA, thereby inhibiting the replication of cancer cell DNA and...

Embodiment 2

[0029] The preparation method of the tuna extract with the effect of inhibiting cancer cells comprises the following steps:

[0030] 1) Compound enzyme enzymatic hydrolysis: use Brevibacterium oxidans to produce alkaline protease and wakame protease to enzymolyze tuna fish meat homogenate, and use 28kHZ ultrasonic to assist enzymolysis. The amount of alkaline protease and wakame protease produced by Brevibacterium oxidans was 1.9wt%, the enzymolysis temperature was 32°C, the pH was 7.8, and the enzymolysis time was 65min;

[0031] 2) Isoelectric point precipitation: adjust the pH of the enzymolysis solution to 6.2 with 1mol / L dilute hydrochloric acid, and centrifuge to collect the precipitate after standing for 2 hours;

[0032] 3) Ultrafiltration and nanofiltration: Dissolve the precipitate obtained by isoelectric point precipitation with 2 times the volume of pure water, first use a 3kDa ultrafiltration membrane to pressurize ultrafiltration, and then use nanofiltration memb...

Embodiment 3

[0036] The preparation method of the tuna extract with the effect of inhibiting cancer cells comprises the following steps:

[0037] 1) Compound enzyme enzymatic hydrolysis: use Brevibacterium oxidans to produce alkaline protease and wakame protease to enzymolyze tuna fish meat homogenate, and use 26kHZ ultrasonic to assist enzymolysis. The addition amount of Brevibacterium oxidans alkaline protease and wakame protease is 1.8wt%, the mass ratio of Brevibacterium oxidans alkaline protease and wakame protease is 1:2, the enzymatic hydrolysis temperature is 30°C, and the pH is 8.0 , the enzymatic hydrolysis time is 55min;

[0038] 2) Isoelectric point precipitation: adjust the pH of the enzymolysis solution to 6.2 with 1mol / L dilute hydrochloric acid, and centrifuge to collect the precipitate after standing for 2 hours;

[0039] 3) Ultrafiltration and nanofiltration: Dissolve the precipitate obtained by isoelectric point precipitation with 2 times the volume of pure water, first...

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PUM

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Abstract

The invention discloses a preparation method of tuna extract with inhibitory effect on cancer cells, and preparation steps include compound enzyme enzymolysis, isoelectric point precipitation, ultrafiltration, nanofiltration, HPLC elution and freezing drying. The beneficial effects are that tuna protein has a lot of marine Brevibacterium oxydan secreting alkaline protease and undariapinnatifida protease cleavage sites, after the enzymolysis, lots of different lengths of peptide chains can be obtained, and the enzymolysis efficiency is high; the preparation method has the advantages of being simple, less in equipment, low in preparation cost, accurate in operation data of each step, good in repeatability, and suitable for mass production; the purity of the prepared bioactive short peptide is high, DNA double helix structure unwinding of the cancer cells can be effectively inhibited by the short peptides, so that the DNA replication of the cancer cells can be inhibited, the cancer cells stay in interphase of cell division, secretion of E-cadherin can be promoted, moveability of the cancer cells is reduced, and the transfer speed of the cancer cells is reduced.

Description

technical field [0001] The invention relates to the technical field of preparation of health care products, in particular to a preparation method of tuna extract which has the function of inhibiting cancer cells. Background technique [0002] As we all know, malignant tumor has become a major disease that threatens human health, and its treatment has also become one of the hot spots that people pay attention to. Traditional chemotherapy and radiotherapy also produce serious side effects during treatment. Tumor biotherapy is the fourth treatment after traditional therapy. Current research has confirmed that biotherapy is playing an increasingly important role in the treatment of tumors and is expected to become a therapy for overcoming tumors. [0003] Peptides have only been paid attention to since the beginning of the 20th century. Studies have found that they are different from proteins and amino acids, and have the effect of drugs. It was not until the 1990s that there ...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/16C07K1/30C07K1/34
CPCC12P21/06C07K1/16C07K1/30C07K1/34
Inventor 刘洋
Owner 浦江县泰如食品科技有限公司
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