CN1814775A - Method for preparing antiviral active oligomeric peptide - Google Patents
Method for preparing antiviral active oligomeric peptide Download PDFInfo
- Publication number
- CN1814775A CN1814775A CN 200510045516 CN200510045516A CN1814775A CN 1814775 A CN1814775 A CN 1814775A CN 200510045516 CN200510045516 CN 200510045516 CN 200510045516 A CN200510045516 A CN 200510045516A CN 1814775 A CN1814775 A CN 1814775A
- Authority
- CN
- China
- Prior art keywords
- meat
- oyster
- constant temperature
- sephadex
- oligomeric peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
This invention relates to a method for preparing antivirus active oligomerized peptides by step directional enzymolyzing oysters with a compound enzyme characterizing in adding Tris-HCL buffer solution into the oyster meat to be beated, the increased number of the buffer is 2-3 times of kilograms of the meat, trypsase is added according to the proportion of 60-65U/g meat to be mixed under constant temperature and enzymolyzed for 3-4 hours under 40-50deg.C then to eliminate the enzymes and cool, regulating pH value to 5-6 with an acid and adding pineappleproteinase in the meat proportion of 70-75U/g to be mixed and enzymolyzed for 5-6 hours under the 45-55deg.C constant temperature to eliminate the enzymes and cool, centrifuge to take the supernatant fluid to pass through super-filtration, Sephadex G-25 gel chromaphoresis, DEAE Sephadex A-25 ionic exchange post chromaphoresis and counter-phase high efficient liquid phase chromqtogram separation and purification to be frozen and dried.
Description
Technical field
The present invention relates to a kind ofly produce method with antiviral active oligomeric peptide with prozyme directional enzymatic oyster.
Background technology
Virus is a class and the closely-related parasitic microbe of human health, in recent years, the appearance of virus such as bird flu, foot and mouth disease, mad cow disease, severe acute respiratory syndrome, acquired immune deficiency syndrome (AIDS), monkey smallpox, Xi Niluo and Ebola and popular, human beings'health in serious threat.The disease that virus causes not only has highly pathogenic, and has the intensive infectivity.Therefore, the prevention of these diseases is human great, a persistent difficult problem that is faced with treatment.
From nineteen forty-one penicillin come out, antibiotic fast development is controlled effectively many bacterial infection diseases, it is then much more difficult than the former to prevent and treat virus disease, has become the outstanding problem of present communicable disease.Because the structure and the propagation mode of virus are different from bacterium, they lack the enzyme system of the breeding of self, must colonize in the host cell, the nucleic acid and the protein ability growth and breeding that synthesize himself by means of the enzyme system of host cell, this just makes medicine also can kill and wound host's normal cell in to virus generation effect, so make the application of antiviral drug be subjected to certain restriction.In addition, the clinical symptom of virus infection occurs after the peak of the viral growth of being everlasting, and also causes medicine to be difficult to play a role.
At present, the medicine of treatment virus disease mostly is Western medicine greatly, has all side effects.Do not see the report of the antiviral oligomeric peptide of preparation from oyster albumen as yet.
Summary of the invention
The purpose of this invention is to provide and a kind ofly produce the method with antiviral active oligomeric peptide with prozyme substep directional enzymatic oyster, it can remedy the above-mentioned deficiency of prior art.
A kind of method of producing oligomeric peptide with prozyme substep directional enzymatic oyster with antiviral activity, it is characterized in that adding the Tris-HCl damping fluid to Oyster, making beating, this damping fluid rise number be Oyster kilogram number 2-3 doubly, ratio according to 60-65U/g meat adds trypsinase, and constant temperature stirred, at 40-50 ℃ of following enzymolysis 3-4 hour, the enzyme that goes out then, cooling; Transfer pH to 5-6 with acid, ratio according to 70-75U/g meat adds bromeline, stir, 45-55 ℃ of following constant temperature enzymolysis 5-6 hour, the enzyme that goes out, cooling, centrifugal, after getting the ultrafiltration of supernatant liquor process, Sephadex G-25 gel filtration chromatography, DEAE Sephadex A-25 ion-exchange chromatography, RPLC separation and purification, freeze-drying.
The present invention is a raw material with the oyster of low value, utilizes the preparation of trypsinase and bromeline complex enzyme hydrolysis to have the oligomeric peptide of antiviral activity, and products therefrom is active high, and influenza virus and simplexvirus are had good inhibitory effect.
Description of drawings
Accompanying drawing is the inhibition design sketch of active oligomeric peptide of the present invention to virus.A is the cell contrast among the figure, and B is a virus control, and C is the 1# sample, and D is the 2# sample.
Embodiment
Add the Tris-HCl damping fluid of 300ml to 100 gram Oysters, making beating, gained slurries pH is 7.5-8.5, preferably 8.0.According to 60-65U/g meat, preferably the ratio of 60U/g meat adds trypsinase, and constant temperature stirs, under 40-50 ℃, preferably 50 ℃ following enzymolysis 3-4 hour, preferably 3 hours, the 95-100 ℃ of enzyme that goes out was cooled to 45-55 ℃ then, best 50 ℃; Transfer pH to 5-6 with hydrochloric acid, preferably 5.5, according to 70-75U/g meat, preferably the ratio of 700U/g meat adds bromeline, stir, at 45-55 ℃, preferably 50 ℃ of following constant temperature enzymolysis are 5-6 hour, and preferably 5 hours, 95-100 ℃ of enzyme that goes out then, be cooled to normal temperature, frozen centrifugation under the rotating speed of 9000-10000r/min is got the supernatant liquor ultrafiltration, collects the part of molecular weight 5-10K, again through Sephadex G-25 gel filtration chromatography, DEAE Sephadex A-25 ion-exchange chromatography, RPLC separation and purification, freeze-drying promptly obtain having the peptide of antiviral activity.
The concentration of used Tris-HCl damping fluid is 0.04-0.06.M among the present invention, and pH is 7.5-8.5, its consumption rise number be Oyster kilogram number 2-3 doubly.
Adopt viral microneutralization laboratory method that the anti-influenza virus activity of 1# (oligomeric peptide of molecular weight 5-10k) and 2# (oligomeric peptide of molecular weight 1-5k) sample is detected, the result as shown in drawings.Experimental result shows that with virus control relatively, 1# sample (being the peptide of the alleged molecular weight of the present invention at 5-10K) has certain inhibition effect to influenza virus, is kept perfectly by the part cell after 72 hours at virus infection; After 96 hours, the complete pathology of cell.
Claims (1)
1, a kind of method of producing oligomeric peptide with prozyme substep directional enzymatic oyster with antiviral activity, it is characterized in that adding the Tris-HCl damping fluid to Oyster, making beating, this damping fluid rise number be Oyster kilogram number 2-3 doubly, ratio according to 60-65U/g meat adds trypsinase, and constant temperature stirred, at 40-50 ℃ of following enzymolysis 3-4 hour, the enzyme that goes out then, cooling; Transfer pH to 5-6 with acid, ratio according to 70-75U/g meat adds bromeline, stir, 45-55 ℃ of following constant temperature enzymolysis 5-6 hour, the enzyme that goes out, cooling, centrifugal, after getting the ultrafiltration of supernatant liquor process, Sephadex G-25 gel filtration chromatography, DEAE SephadexA-25 ion-exchange chromatography, RPLC separation and purification, freeze-drying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510045516 CN1814775A (en) | 2005-12-06 | 2005-12-06 | Method for preparing antiviral active oligomeric peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510045516 CN1814775A (en) | 2005-12-06 | 2005-12-06 | Method for preparing antiviral active oligomeric peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1814775A true CN1814775A (en) | 2006-08-09 |
Family
ID=36907131
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200510045516 Pending CN1814775A (en) | 2005-12-06 | 2005-12-06 | Method for preparing antiviral active oligomeric peptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1814775A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1908183B (en) * | 2006-08-16 | 2010-09-29 | 上海奥利实业有限公司 | Enzymolytic hybrid peptide of native protein with endothelin antagonism |
| CN103263382A (en) * | 2013-05-08 | 2013-08-28 | 山东大学（威海） | Oyster polysaccharide gel formulation capable of playing protection role in immunological liver injury |
| CN104039808A (en) * | 2011-11-23 | 2014-09-10 | 赛诺菲 | Protein purification using BIS-TRIS buffer |
| US10793622B2 (en) | 2013-05-06 | 2020-10-06 | Sanofi | Continuous multistep process for purifying antibodies |
-
2005
- 2005-12-06 CN CN 200510045516 patent/CN1814775A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1908183B (en) * | 2006-08-16 | 2010-09-29 | 上海奥利实业有限公司 | Enzymolytic hybrid peptide of native protein with endothelin antagonism |
| CN104039808A (en) * | 2011-11-23 | 2014-09-10 | 赛诺菲 | Protein purification using BIS-TRIS buffer |
| US10131714B2 (en) | 2011-11-23 | 2018-11-20 | Sanofi | Protein purification using bis-tris buffer |
| US10793622B2 (en) | 2013-05-06 | 2020-10-06 | Sanofi | Continuous multistep process for purifying antibodies |
| CN103263382A (en) * | 2013-05-08 | 2013-08-28 | 山东大学（威海） | Oyster polysaccharide gel formulation capable of playing protection role in immunological liver injury |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20150111247A1 (en) | Method of double-coating lactic acid bacteria | |
| CN1330771C (en) | Method of preparing peptide having trypsin inhibition activity by using complex enzyme enzymolysis oyster | |
| US20100113383A1 (en) | Bifidobacterial gene sequences and their use | |
| CN110809411A (en) | Composition comprising lactic acid bacteria having improved colonization in intestinal tract by coating silk fibroin | |
| KR100429495B1 (en) | Manufacturing method of Dual-coated Lactic acid bacteria powder using protein and polysaccharlde | |
| CN105209610A (en) | Novel bacteriophage and antibacterial composition comprising the same | |
| CN1814775A (en) | Method for preparing antiviral active oligomeric peptide | |
| KR20190019087A (en) | Compositions and methods for total microbial therapy | |
| CN105324481A (en) | Novel bacteriophage and antibacterial composition comprising the same | |
| CN110157645A (en) | A kind of Lactobacillus salivarius Y4 and its application | |
| CN110527704A (en) | A method of synthesis cream-N- disaccharides | |
| CN115960167A (en) | Corn anti-adhesion peptide and preparation method and application thereof | |
| CN103734745B (en) | Method for preparing compound microcapsule of high-concentration lactoferrin and lactobacillus acidophilus | |
| KR100429494B1 (en) | Manufacturing method of Protein- coated Lactic acid bacteria powder | |
| CN114214366A (en) | Compound medicine of small peptide powder and heme peptide red for preventing and treating anemia and preparation method and application thereof | |
| WO2010044688A1 (en) | A method of producing peptide preparations for oral administration | |
| WO2017112945A1 (en) | Compositions and methods of use of novel strains of lactobacillus fermentum | |
| CN109837219B (en) | Separation and purification of hydrolyzed cow milk allergen beta-lactoglobulin protease and application thereof | |
| CN104087642A (en) | Method for making protein polypeptide | |
| CN109820088B (en) | Wheat germ protein hydrolysate and preparation method and application thereof | |
| CN107974443A (en) | A kind of acylated protein enzymatic compositions for being used to prepare spleen peptide | |
| CN103549435A (en) | Method for preparing oral whey protein peptide liquid | |
| CN103621762A (en) | Method for preparing low-molecular-weight oyster peptide by employing bacillus natto fermentation process | |
| CN107058266B (en) | A method of lysozyme is prepared by zymotechnique | |
| JP2001240599A (en) | Lactic oligosaccharide fraction |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |