CN101519442A - Method for inducing silkworms to produce antibacterial peptides - Google Patents
Method for inducing silkworms to produce antibacterial peptides Download PDFInfo
- Publication number
- CN101519442A CN101519442A CN200910116463A CN200910116463A CN101519442A CN 101519442 A CN101519442 A CN 101519442A CN 200910116463 A CN200910116463 A CN 200910116463A CN 200910116463 A CN200910116463 A CN 200910116463A CN 101519442 A CN101519442 A CN 101519442A
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- CN
- China
- Prior art keywords
- silkworms
- antibacterial peptides
- silkworm
- produce antibacterial
- inducing
- 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.)
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Links
- 241000255789 Bombyx mori Species 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 31
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 19
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 18
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 18
- 230000001939 inductive effect Effects 0.000 title claims abstract description 17
- 241000700605 Viruses Species 0.000 claims abstract description 10
- 241000894006 Bacteria Species 0.000 claims abstract description 9
- 235000008708 Morus alba Nutrition 0.000 claims abstract description 6
- 240000000249 Morus alba Species 0.000 claims abstract description 6
- 239000003910 polypeptide antibiotic agent Substances 0.000 claims description 24
- 102000044503 Antimicrobial Peptides Human genes 0.000 claims description 11
- 108700042778 Antimicrobial Peptides Proteins 0.000 claims description 11
- 238000007710 freezing Methods 0.000 claims description 11
- 230000008014 freezing Effects 0.000 claims description 11
- 238000004321 preservation Methods 0.000 claims description 10
- 241000607142 Salmonella Species 0.000 claims description 3
- 235000003642 hunger Nutrition 0.000 claims description 3
- 230000000968 intestinal effect Effects 0.000 claims description 2
- 238000000605 extraction Methods 0.000 abstract description 4
- 238000002347 injection Methods 0.000 abstract description 3
- 239000007924 injection Substances 0.000 abstract description 3
- 239000006228 supernatant Substances 0.000 description 5
- 230000006698 induction Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000361919 Metaphire sieboldi Species 0.000 description 1
- 241000257159 Musca domestica Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000001857 anti-mycotic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000005360 mashing Methods 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 231100000255 pathogenic effect Toxicity 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000037351 starvation Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Fodder In General (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to a method for inducing silkworms to produce antibacterial peptides. In the method, a virus or bacterium injection method can be adopted or a method enabling silkworms to be hungry can be adopted. Micro bacteria or viruses are injected to the silkworms when adopting the virus or bacterium injection method. When the method enabling silkworms to be hungry is adopted, no mulberry leaf is fed to the silkworms which are hungry for 48 hours and then normally fed for 24 hours. The method for inducing silkworms to produce antibacterial peptides is simple and easy to carry out, can effectively induce more antibacterial peptides and enables the extraction efficiency of the antibacterial peptides to be greatly improved.
Description
Technical field
The present invention relates to a kind of induction method of antibacterial peptide, especially a kind of induction method of cultivated silkworm antimicrobial peptide.
Background technology
Antibacterial peptide is the defensive polypeptide active substance that a class of organism immunity defence system generation is resisted exogenous pathogenic agent pathogenic effects, is the important composition composition of organism congenital immunity, has broad spectrum antibiotic activity.Antibacterial peptide also has antiviral, antimycotic and antitumor action except antibacterium, be good natural antibacterial medicament sources, and it is little to have a relative molecular mass, Heat stability is good, non-immunogenicity, characteristics such as antibiotic wide spectrum.Antibacterial peptide still be a kind ofly have no side effect, the environment-friendly type preparation of noresidue, no bacterial drug resistance, have advantages such as thermally-stabilised, good water solubility, broad-spectrum sterilization, and livestock and poultry had the function of promotes growth, health care and treatment disease.Therefore, the application prospect of antibacterial peptide is had an optimistic view of by people.Present the inducing of antibacterial peptide in the technology, the material that uses that has is housefly, the material that uses that has is earthworm.But the getable antibacterial peptide of institute is less, and the inductive difficulty is also bigger.
Summary of the invention
The method that the purpose of this invention is to provide a kind of inducing silkworms to produce antibacterial peptides induces difficulty big to solve present antibacterial peptide, the problem that resultant antibacterial peptide is few.
In order to achieve the above object, the technical solution used in the present invention is:
A kind of method of inducing silkworms to produce antibacterial peptides is characterized in that: carry out following steps successively:
(1) injects bacterium or the virus of micro-0.05ul～0.5ul at the link position of silkworm;
(2) normally raise 12-36 hour after, be put into freezing preservation in-20～-40 ℃ of environment;
(3) take out shred, smash to pieces, homogenate, extract cultivated silkworm antimicrobial peptide wherein.
A kind of method of inducing silkworms to produce antibacterial peptides is characterized in that: described method is for making silkworm hunger, and the step of described method is:
(1) do not feed mulberry leaf to silkworm, allow its hungry 24-48 hour;
(2) normally raise 12-36 hour then after, be put into freezing preservation in-20～-40 ℃ of environment;
(3) take out shred, smash to pieces, homogenate, extract cultivated silkworm antimicrobial peptide wherein.
The method of described inducing silkworms to produce antibacterial peptides is characterized in that: after normally raising 24 hours, be put into freezing preservation in-30 ℃ of environment.
The method of described inducing silkworms to produce antibacterial peptides is characterized in that described bacterium or virus are meant intestinal bacteria, Salmonellas, NPV virus.
The method of described inducing silkworms to produce antibacterial peptides is characterized in that: do not feed mulberry leaf to silkworm, allow its hungry 48 hours; After normally raising 24 hours then, be put into freezing preservation in-30 ℃ of environment.
The induction method of cultivated silkworm antimicrobial peptide involved in the present invention is simple, and is easy to implement, and can produce more antibacterial peptide, improved the extraction efficiency of antibacterial peptide greatly.
Embodiment
Example 1
A kind of production method of cultivated silkworm antimicrobial peptide:
(1) Salmonellas or the NPV virus of injecting micro-0.05ul～0.5ul for silkworm link position;
(2) normally raise 24 hours after, be put into freezing preservation in-30 ℃ of environment;
(3) take out shred, smash to pieces, homogenate, extract cultivated silkworm antimicrobial peptide wherein.
Example 2
A kind of production method of cultivated silkworm antimicrobial peptide:
(1) do not feed mulberry leaf to silkworm, allow its hungry 48 hours;
(2) normally raise 24 hours then after, be put into freezing preservation in-30 ℃ of environment;
(3) take out shred, smash to pieces, homogenate, extract cultivated silkworm antimicrobial peptide wherein.
The Acetic Acid Extraction method is adopted in the extraction of antibacterial peptide.Take that different treatment crosses five age silkworm weigh respectively, freezing, shred and with tissue mashing machine smash to pieces, homogenate.Mix with equal-volume 5% acetate then, stir the back water proof that spends the night under 4 ℃ of conditions and boil 15min, cooling rapidly.Mixture is 8000rpm under 4 ℃ of conditions, and centrifugal 30min gets supernatant liquor, preserves under 4 ℃ of conditions.Throw out mixes with the acetate of equal-volume 5%, spends the night in 4 ℃ of stirrings, lixiviates once more, repeats above-mentioned centrifugal process, get supernatant, merge supernatant liquor twice, adjust the pH value, recentrifuge is abandoned precipitation, and the supernatant liquor lyophilize is the raw product of cultivated silkworm antimicrobial peptide class material.Get supernatant liquor and in another tool plug plastic centrifuge tube, preserve (, can in-20 ℃ refrigerated tank, preserve 30d) down if do not carry out the mensuration of antimicrobial substance immediately in low temperature.
The antibacterial peptide of silkworm being induced generation with different methods is difference slightly
| Induction method | Injection | Starvation method | Normal silkworm |
| Antibacterial circle diameter (mm) | 2.1 | 2.3 | 1.4 |
Claims (5)
1, a kind of method of inducing silkworms to produce antibacterial peptides is characterized in that: carry out following steps successively:
(1) injects bacterium or the virus of micro-0.05ul～0.5ul at the link position of silkworm;
(2) normally raise 12-36 hour after, be put into freezing preservation in-20～-40 ℃ of environment;
(3) take out shred, smash to pieces, homogenate, extract cultivated silkworm antimicrobial peptide wherein.
2, a kind of method of inducing silkworms to produce antibacterial peptides is characterized in that: described method is for making silkworm hunger, and the step of described method is:
(1) do not feed mulberry leaf to silkworm, allow its hungry 24-48 hour;
(2) normally raise 12-36 hour then after, be put into freezing preservation in-20～-40 ℃ of environment;
(3) take out shred, smash to pieces, homogenate, extract cultivated silkworm antimicrobial peptide wherein.
3, the method for inducing silkworms to produce antibacterial peptides according to claim 1 is characterized in that: after normally raising 24 hours, be put into freezing preservation in-30 ℃ of environment.
4, the method for inducing silkworms to produce antibacterial peptides according to claim 1 is characterized in that described bacterium or virus are meant intestinal bacteria, Salmonellas, NPV virus.
5, the method for inducing silkworms to produce antibacterial peptides according to claim 2 is characterized in that: do not feed mulberry leaf to silkworm, allow its hungry 48 hours; After normally raising 24 hours then, be put into freezing preservation in-30 ℃ of environment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101164633A CN101519442B (en) | 2009-03-31 | 2009-03-31 | Method for inducing silkworms to produce antibacterial peptides |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101164633A CN101519442B (en) | 2009-03-31 | 2009-03-31 | Method for inducing silkworms to produce antibacterial peptides |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101519442A true CN101519442A (en) | 2009-09-02 |
| CN101519442B CN101519442B (en) | 2011-09-28 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101164633A Expired - Fee Related CN101519442B (en) | 2009-03-31 | 2009-03-31 | Method for inducing silkworms to produce antibacterial peptides |
Country Status (1)
| Country | Link |
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| CN (1) | CN101519442B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102613249A (en) * | 2012-02-22 | 2012-08-01 | 湛江师范学院 | Process for preparing board-spectrum antibiotics by immune induction biotechnology |
| CN103621564A (en) * | 2013-12-17 | 2014-03-12 | 湛江师范学院 | Preparation method of biological antibacterial agent |
| CN103641631A (en) * | 2013-12-17 | 2014-03-19 | 湛江师范学院 | Broad-spectrum antibacterial biological fertilizer |
| CN103667303A (en) * | 2013-12-04 | 2014-03-26 | 浙江省海洋开发研究院 | Expression method of antibacterial peptide Hclocidin18 in escherichia coli |
| CN104388508A (en) * | 2014-10-24 | 2015-03-04 | 江西省新龙生物科技有限公司 | Method for splitting insect body by utilizing insect virus to extract insect peptide |
| CN110150478A (en) * | 2019-06-28 | 2019-08-23 | 青岛宝创生物科技有限公司 | A kind of feed addictive and the preparation method and application thereof reducing mastitis for milk cows disease incidence |
| CN118047841A (en) * | 2024-04-09 | 2024-05-17 | 中国农业科学院农业基因组研究所 | Fermented mulberry leaf antibacterial peptide Squ and application thereof |
-
2009
- 2009-03-31 CN CN2009101164633A patent/CN101519442B/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102613249A (en) * | 2012-02-22 | 2012-08-01 | 湛江师范学院 | Process for preparing board-spectrum antibiotics by immune induction biotechnology |
| CN103667303A (en) * | 2013-12-04 | 2014-03-26 | 浙江省海洋开发研究院 | Expression method of antibacterial peptide Hclocidin18 in escherichia coli |
| CN103621564A (en) * | 2013-12-17 | 2014-03-12 | 湛江师范学院 | Preparation method of biological antibacterial agent |
| CN103641631A (en) * | 2013-12-17 | 2014-03-19 | 湛江师范学院 | Broad-spectrum antibacterial biological fertilizer |
| CN103641631B (en) * | 2013-12-17 | 2015-05-13 | 湛江师范学院 | Broad-spectrum antibacterial biological fertilizer |
| CN103621564B (en) * | 2013-12-17 | 2016-05-25 | 湛江师范学院 | A kind of preparation method of biological antibiosis agent |
| CN104388508A (en) * | 2014-10-24 | 2015-03-04 | 江西省新龙生物科技有限公司 | Method for splitting insect body by utilizing insect virus to extract insect peptide |
| CN110150478A (en) * | 2019-06-28 | 2019-08-23 | 青岛宝创生物科技有限公司 | A kind of feed addictive and the preparation method and application thereof reducing mastitis for milk cows disease incidence |
| CN118047841A (en) * | 2024-04-09 | 2024-05-17 | 中国农业科学院农业基因组研究所 | Fermented mulberry leaf antibacterial peptide Squ and application thereof |
| CN118047841B (en) * | 2024-04-09 | 2024-06-11 | 中国农业科学院农业基因组研究所 | Fermented mulberry leaf antibacterial peptide Squ and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101519442B (en) | 2011-09-28 |
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| C53 | Correction of patent for invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Fan Tao Inventor after: Wu Penghao Inventor after: Zhang Yuping Inventor after: Ling Yonghua Inventor after: Zhou Yefeng Inventor after: Dai Junjun Inventor after: Fan Houwen Inventor after: Wang Chuyan Inventor after: Xiao Linzhen Inventor after: Wu Chuanhua Inventor after: Wei Guoqing Inventor after: Tian Shanfu Inventor before: Fan Tao Inventor before: Zhou Yefeng Inventor before: Dai Junjun Inventor before: Fan Houwen Inventor before: Wang Chuyan Inventor before: Xiao Linzhen Inventor before: Wu Chuanhua Inventor before: Wei Guoqing Inventor before: Tian Shanfu |
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| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: FAN TAO ZHOU YEFENG DAI JUNJUN FAN HOUWEN WANG CHUYAN XIAO LINZHEN WU ZHUANHUA WEI GUOQING TIAN SHANFU TO: FAN TAO ZHOU YEFENG DAI JUNJUN FAN HOUWEN WANG CHUYAN XIAO LINZHEN WU ZHUANHUA WEI GUOQING TIAN SHANFU WU PENGHAO ZHANG YUPING LING YONGHUA |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Fan Tao Inventor after: Tian Shanfu Inventor after: Wu Penghao Inventor after: Zhang Yuping Inventor after: Ling Yonghua Inventor after: Zhou Yefeng Inventor after: Dai Junjun Inventor after: Wang Yuting Inventor after: Fan Houwen Inventor after: Wang Chuyan Inventor after: Xiao Linzhen Inventor after: Wu Chuanhua Inventor after: Wei Guoqing Inventor before: Fan Tao Inventor before: Wu Penghao Inventor before: Zhang Yuping Inventor before: Ling Yonghua Inventor before: Zhou Yefeng Inventor before: Dai Junjun Inventor before: Fan Houwen Inventor before: Wang Chuyan Inventor before: Xiao Linzhen Inventor before: Wu Chuanhua Inventor before: Wei Guoqing Inventor before: Tian Shanfu |
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| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110928 Termination date: 20180331 |