CN100432106C - Method for conserving antiseptic peptide using attachment method - Google Patents
Method for conserving antiseptic peptide using attachment method Download PDFInfo
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- CN100432106C CN100432106C CNB2006100357599A CN200610035759A CN100432106C CN 100432106 C CN100432106 C CN 100432106C CN B2006100357599 A CNB2006100357599 A CN B2006100357599A CN 200610035759 A CN200610035759 A CN 200610035759A CN 100432106 C CN100432106 C CN 100432106C
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- antibacterial peptide
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Abstract
The present invention provides a method for preserving antibacterial peptide by an attaching method. The present invention has the technical schemes that an antibacterial peptide solution is immersed in a carrier of absorbent paper or fibre fabrics; then, the carrier containing antibacterial peptide components is dried; the carrier is packaged in vacuum and is preserved below 30 DEG C. The activity maintenance time of the antibacterial peptide preserved by the method of the present invention is obviously longer than that of a preservation method by adding inhibiting agents, and is equivalent to that of preserved samples by a freeze-drying method. Compared with the preservation method by adding inhibiting agents and the freeze-drying method, the method for preserving antibacterial peptide by an attaching method has the advantages of low expense, convenient use and no toxic and side effect.
Description
Technical field
The present invention relates to microbial technology field, specifically relate to a kind of antibacterial peptide method for preserving.
Background technology
Present traditional antibiotic resistance situation is increasingly serious, human beings'health has been constituted serious threat, in order to solve the resistance problem, scientific circles have found antibacterial peptide, the superiority that antibacterial peptide shows is to be difficult for developing immunity to drugs, and the noresidue phenomenon simultaneously, also shows active preferably to protozoon, cancer cells and virus when antibacterial peptide shows broad spectrum antibiotic activity.However but the popularization and application of antibacterial peptide is also premature yet, this mainly is because at present under antibacterial peptide activity and natural antibacterial peptide extracted amount condition of limited, prepare traditional antibiotic substitute, and its cost is very high; Another important factor is its preservation problem, compares with traditional microbiotic, and antibacterial peptide loses activity because of oxidation or enzymolysis easily.Mainly be to add inhibitor and two kinds of approach of freeze-drying preservation aspect the antibacterial peptide preservation at present, there is side effect in the former in these two approach, the popularization and application that is not only restricting as medicine also produces certain interference to scientific effort simultaneously, and latter's cost is higher and be unfavorable for popular operation, therefore in order to promote the popularization and application of antibacterial peptide, need in preservation, to open up novel method.
Summary of the invention
The object of the invention is to provide a kind of easy to use and antibacterial peptide method for preserving of having no side effect.
Antibacterial peptide method for preserving provided by the present invention, its technical scheme is: antibacterial peptide solution is immersed in the carrier of thieving paper or fabric, will contains the carrier oven dry of antibacterial peptide composition then, carry out vacuum packaging again, in preservation below 30 ℃.
Said thieving paper carrier preferably adopts filter paper.
Said fabric carrier can be hospital gauze, also comprises non-woven fabrics.
Further scheme is, the carrier that will contain the antibacterial peptide composition carries out vacuum packaging and in 4 ℃ of preservations.
For the strong antibacterial peptide preservation as stated above again after 100 ℃ of thermal treatment of thermotolerance, then stability is stronger.
The specific operation process of the inventive method is as follows:
1. filter paper and gauze are cut into wide 0.5cm respectively, the rectangular back sterilizing, drying of long 2cm is also preserved standby in drying at room temperature.
2. dry after antibacterial peptide solution being immersed carrier.
3. the carrier that will contain the antibacterial peptide composition carries out vacuum packaging and in preservation below 30 ℃, is preferably in 4 ℃ of preservations.
The antibacterial peptide of the antibacterial peptide of the inventive method preservation and existing method for preserving preservation compares:
Wherein, the method for existing preservation antibacterial peptide comprises:
One, adds the method for preserving of inhibitor
In antibacterial peptide solution, add behind the inhibitor (phenylthiourea 20 μ g/ml, PMSF30 μ g/ml and 20 μ g/mlAprotinin) directly in-80 ℃ of preservations.
Two, lyophilization preservation
Antibacterial peptide solution is divided in the centrifuge tube of 1.5mL, puts-80 ℃ of quick-frozens, put into Freeze Drying Equipment then and be lyophilized into powdery, change-20 ℃ of preservations over to.
The antibacterial peptide of the antibacterial peptide of the inventive method preservation and existing method for preserving preservation is active to be detected relatively by following method, and the result is as follows:
1. solid LB substratum is melted, treat that temperature adds the test bacterium when reducing to about 45 ℃ (non-scald on hand) and shakes up (making substratum contain the about 1 * 106cells/ μ of the concentration L of bacterium), pour culture dish then into.
2. the sample of liquid towards utilizes agarose hole diffusion process to detect its anti-microbial activity, the sample of preserving with lyophilized powder needs with aseptic ddH20 heavy molten before doing bacteriostatic experiment, the bacteriostatic test method is used agarose hole diffusion process equally, then carrier on average is cut into directly to be affixed on behind 4 equal portions for the sample of preservation of the present invention and contains bacterium culture medium surface (every part of contained antibacterial peptide amount is identical with added amount in the diffusion process mesopore of agarose hole), put 37 ℃ of overnight incubation, measure antibacterial circle diameter.
3. active detected respectively on same day preservation day and carry out when playing 5 days, 10 days, 20 days, 30 days, 40 days, 50 days, 60 days preservation day.
Experimental result (seeing table 1 and table 2 in the accompanying drawing): table 1 is the antibacterial result of several preservation samples of bollworm antibacterial peptide, shows in the table that the reduced activity of inhibitor group is the most obvious, and activity has weakened more than half after 60 days; The active amount of decrease of the sample of freeze-drying group and two kinds of attachment method preservations activity slowly and after 60 days still is retained in more than 50%.Table 2 is the antibacterial result of several preservation samples of antibacterial peptide Hex-Mag, and the result shows that the reduced activity of inhibitor group is the fastest, and the active amount of decrease of the sample of freeze-drying group and two kinds of attachment method preservations slowly and can both keep activity more than 50% after 60 days.
Table 1: the bollworm antibacterial peptide of different condition preservation is in the bacteriostatic experiment result of different times
(antibacterial circle diameter: millimeter)
Table 2: the antibacterial peptide Hex-Mag of different condition preservation is in the bacteriostatic experiment result of different times
(antibacterial circle diameter: millimeter)
The result proves the antibacterial peptide of the inventive method institute preservation, the time ratio that its activity is kept adds the method for preserving of inhibitor and obviously will grow, hold time quite with the sample activity of lyophilization preservation, and compare with these two kinds of methods, the inventive method preservation antibacterial peptide expense is low, conveniently uses and has no side effect.
Embodiment
Embodiment one
The preservation of bollworm antibacterial peptide
1. filter paper is cut into wide 0.5cm, the rectangular back sterilizing, drying of long 2cm is also preserved standby in drying at room temperature.
2. dry after bollworm antibacterial peptide solution being immersed the filter paper carrier.
3. the filter paper carrier that will contain bollworm antibacterial peptide composition carries out vacuum packaging and in 4 ℃ of preservations.
The result shows, the active amount of decrease of the sample of this attachment method preservation activity slowly and after 60 days still is retained in more than 50%.
Embodiment two
The preservation of antibacterial peptide Hex-Mag
1. hospital gauze is cut into wide 0.5cm, the rectangular back sterilizing, drying of long 2cm is also preserved standby in drying at room temperature.
2. dry after antibacterial peptide Hex-Mag solution being immersed the hospital gauze carrier.
3. the hospital gauze carrier that will contain antibacterial peptide Hex-Mag composition carries out vacuum packaging and in 4 ℃ of preservations.
The result shows, the active amount of decrease of the sample of this attachment method preservation slowly and can both keep activity more than 50% after 60 days.
Claims (3)
1. the method for an attachment method preservation antibacterial peptide is characterized in that antibacterial peptide solution is immersed in the carrier of thieving paper after the sterilization or hospital gauze or non-woven fabrics, will contain the carrier oven dry of antibacterial peptide composition then, carries out vacuum packaging again, in preservation below 4 ℃.
2. the method for attachment method preservation antibacterial peptide according to claim 1 is characterized in that said thieving paper carrier is a filter paper.
3. the method for attachment method preservation antibacterial peptide according to claim 1, it is characterized in that will preservation antibacterial peptide solution after 100 ℃ of following thermal treatment, again by the described method preservation of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100357599A CN100432106C (en) | 2006-06-02 | 2006-06-02 | Method for conserving antiseptic peptide using attachment method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100357599A CN100432106C (en) | 2006-06-02 | 2006-06-02 | Method for conserving antiseptic peptide using attachment method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1884298A CN1884298A (en) | 2006-12-27 |
| CN100432106C true CN100432106C (en) | 2008-11-12 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006100357599A Expired - Fee Related CN100432106C (en) | 2006-06-02 | 2006-06-02 | Method for conserving antiseptic peptide using attachment method |
Country Status (1)
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| CN (1) | CN100432106C (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1110323A (en) * | 1993-10-14 | 1995-10-18 | 小野药品工业株式会社 | Novel polypeptides and DNAS encoding them |
| CN1216768A (en) * | 1998-10-29 | 1999-05-19 | 吉林大学 | Biologic antibiotic peptide, and method for preparing same |
| US6630583B1 (en) * | 1998-05-06 | 2003-10-07 | St. Jude Children's Research Hospital | Antibiotics and methods of using the same |
-
2006
- 2006-06-02 CN CNB2006100357599A patent/CN100432106C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1110323A (en) * | 1993-10-14 | 1995-10-18 | 小野药品工业株式会社 | Novel polypeptides and DNAS encoding them |
| US6630583B1 (en) * | 1998-05-06 | 2003-10-07 | St. Jude Children's Research Hospital | Antibiotics and methods of using the same |
| CN1216768A (en) * | 1998-10-29 | 1999-05-19 | 吉林大学 | Biologic antibiotic peptide, and method for preparing same |
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| Publication number | Publication date |
|---|---|
| CN1884298A (en) | 2006-12-27 |
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Granted publication date: 20081112 Termination date: 20130602 |