CN112250884A - Dendritic polymeric antibacterial peptide and preparation method and application thereof - Google Patents
Dendritic polymeric antibacterial peptide and preparation method and application thereof Download PDFInfo
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- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims abstract description 99
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 41
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 37
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 33
- 239000000412 dendrimer Substances 0.000 claims abstract description 21
- 229920000736 dendritic polymer Polymers 0.000 claims abstract description 21
- 244000144972 livestock Species 0.000 claims abstract description 15
- 239000003674 animal food additive Substances 0.000 claims abstract description 10
- 229920001184 polypeptide Polymers 0.000 claims abstract description 10
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- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 6
- 230000000091 immunopotentiator Effects 0.000 claims abstract description 4
- 238000000108 ultra-filtration Methods 0.000 claims description 20
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- 238000000034 method Methods 0.000 claims description 7
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- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 1
- PJWOOBTYQNNRBF-BZSNNMDCSA-N Leu-Phe-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)O)N PJWOOBTYQNNRBF-BZSNNMDCSA-N 0.000 description 1
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- WALVCOOOKULCQM-ULQDDVLXSA-N Lys-Arg-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WALVCOOOKULCQM-ULQDDVLXSA-N 0.000 description 1
- IMAKMJCBYCSMHM-AVGNSLFASA-N Lys-Glu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN IMAKMJCBYCSMHM-AVGNSLFASA-N 0.000 description 1
- LNMKRJJLEFASGA-BZSNNMDCSA-N Lys-Phe-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LNMKRJJLEFASGA-BZSNNMDCSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- PEFJUUYFEGBXFA-BZSNNMDCSA-N Phe-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 PEFJUUYFEGBXFA-BZSNNMDCSA-N 0.000 description 1
- ZSKJPKFTPQCPIH-RCWTZXSCSA-N Pro-Arg-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZSKJPKFTPQCPIH-RCWTZXSCSA-N 0.000 description 1
- 241000607683 Salmonella enterica subsp. enterica serovar Pullorum Species 0.000 description 1
- WFENBJPLZMPVAX-XVKPBYJWSA-N Val-Gly-Glu Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O WFENBJPLZMPVAX-XVKPBYJWSA-N 0.000 description 1
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- -1 amino, carboxyl Chemical group 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/002—Dendritic macromolecules
- C08G83/003—Dendrimers
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G73/00—Macromolecular compounds obtained by reactions forming a linkage containing nitrogen with or without oxygen or carbon in the main chain of the macromolecule, not provided for in groups C08G12/00 - C08G71/00
- C08G73/02—Polyamines
- C08G73/028—Polyamidoamines
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- Polymers & Plastics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Birds (AREA)
- Animal Husbandry (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Polyamides (AREA)
Abstract
The invention discloses a dendritic polymeric antibacterial peptide, a preparation method and application thereof, and relates to the technical field of medical high polymer materials. The antibacterial polypeptide consists of two parts, wherein the first part is polyamide-amine type dendrimer, and the second part is antibacterial peptide. The invention connects the antibacterial peptide with the amino or carboxyl at the tail end of the polyamide-amine type dendrimer by a covalent bonding mode to prepare the dendritic polymeric antibacterial peptide. Finally, the invention provides an application of the poly antibacterial peptide in preparing livestock and poultry feed additives or immunopotentiators. The dendritic antibacterial peptide has broad-spectrum antibacterial performance and good lytic antibacterial activity on gram-negative bacteria and gram-positive bacteria. Compared with the traditional original antibacterial peptide, the poly antibacterial peptide has more action sites and greatly improves the antibacterial activity.
Description
Technical Field
The invention relates to the technical field of medical high polymer materials, in particular to dendritic polymeric antibacterial peptide and a preparation method and application thereof.
Background
With the wide application and even abuse of antibiotics, drug-resistant strains (including multi-drug-resistant strains) are frequently found, the therapeutic effect of the antibiotics is severely challenged, and the development of novel antibacterial products becomes an urgent social demand. Natural antimicrobial peptides are considered to be one of the most promising antibiotic substitutes due to their antimicrobial properties and lack of drug resistance. Although the antibacterial peptide has remarkable advantages in bacteriostasis, if the antibacterial peptide is really a commercial drug, a plurality of problems need to be solved, such as low activity of natural antibacterial peptide and incapability of being directly used as an antibacterial drug.
Disclosure of Invention
In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is to provide a dendritic polymeric antibacterial peptide, a preparation method and an application thereof, such that the dendritic polymeric antibacterial peptide has low cost, is easily metabolized by human bodies or other organisms, has low cytotoxicity, and has a wide application prospect in biomedical applications and the like.
In order to achieve the above object, the present invention provides a dendritic antibacterial polypeptide, wherein the antibacterial polypeptide has a chemical formula: DAm，
Wherein D represents a polyamidoamine-type dendrimer, A represents an antibacterial peptide, and m represents the number of the antibacterial peptides.
The invention provides a preparation method of dendritic polymeric antibacterial peptide, which comprises the following steps:
s100, reacting polyamide-amine type dendritic macromolecules with macromolecules at the tail ends thereof as amino groups with glutaraldehyde to obtain glutaraldehyde-activated polyamide-amine type tree molecules;
s200, removing redundant glutaraldehyde through ultrafiltration, then reacting the polyamide-amine type tree molecules with antibacterial peptide for a certain time, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
The invention also provides a preparation method of the antibacterial polypeptide, which comprises the following steps:
s1000, reacting the polyamide-amine dendrimer of which the tail end is carboxyl and carboxylate with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide to obtain active ester of the polyamide-amine dendrimer;
s2000, removing redundant 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide through ultrafiltration, reacting the active ester with the antibacterial peptide for a certain time, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
Finally, the invention provides an application of the poly antibacterial peptide in preparing livestock and poultry feed additives or immunopotentiators.
Compared with the prior art, the invention has the advantages that:
(1) the invention introduces antibacterial peptide into the tail end of polyamide-amine type dendritic macromolecule to obtain the poly antibacterial peptide, and the poly antibacterial peptide has more action sites compared with the original antibacterial peptide, and the antibacterial performance is improved by 10 times;
(2) the invention can be used as livestock and poultry feed additive or livestock and poultry disease prevention and treatment preparation.
The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings to fully understand the objects, the features and the effects of the present invention.
Drawings
FIG. 1 is a schematic diagram of a process for preparing a poly-antibacterial peptide;
FIG. 2 is a gel electrophoresis image of the poly-antibacterial peptide and the antibacterial peptide;
FIGS. 3(a) and 3(b) are the antibacterial peptides and the poly antibacterial peptides in Escherichia coli C600Comparison of spotting results on dishes, wherein FIG. 3(a) shows the antimicrobial peptide Escherichia coli C600The results of spotting on a dish are shown in FIG. 3(b) for E.coli C600Spotting results on a plate;
FIGS. 4(a) and 4(b) are a comparison of results of deposition of antibacterial peptides and poly-antibacterial peptides on a Salmonella 13076 plate, wherein FIG. 4(a) is a result of deposition of antibacterial peptides on a Salmonella 13076 plate, and FIG. 4(b) is a result of deposition of poly-antibacterial peptides on a Salmonella 13076 plate;
fig. 5(a) and 5(b) are results of spotting of antibacterial peptides and poly antibacterial peptides on a staphylococcus aureus 26001 plate, wherein fig. 5(a) is a result of spotting of antibacterial peptides on a staphylococcus aureus 26001 plate, and fig. 5(b) is a result of spotting of poly antibacterial peptides on a staphylococcus aureus 26001 plate.
Detailed Description
The technical contents of the preferred embodiments of the present invention will be more clearly and easily understood by referring to fig. 1 to 5(b) of the specification. The present invention may be embodied in many different forms of embodiments and the scope of the invention is not limited to the embodiments set forth herein.
The invention provides a dendritic antibacterial peptide, and figure 1 shows the preparation of antibacterial peptideThe process schematic diagram shows the basic preparation principle of the antibacterial polypeptide, and the chemical formula of the antibacterial polypeptide is as follows: DAm
Wherein D represents a polyamidoamine-type dendrimer, A represents an antibacterial peptide, and m represents the number of the antibacterial peptides. It will be appreciated that it is for this reason that the invention calls the resulting product a polypeptide.
The polyamide-amine type dendrimer terminal group can be any one of the following groups: amino, is carboxyl, carboxylate.
The polyamide-amine dendritic macromolecule is a product of 1-3 generations, and mainly utilizes a plurality of functional groups (amino, carboxyl or carboxylate) on the surface of the dendritic macromolecule, for example, the number of the first generation surface is 8, the number of the second generation is 16, the number of the third generation is 32, the number of generations is more, the number of the functional groups is more, but the price of the algebraic high reagent is correspondingly more expensive, the price is very high after the 4 th generation, and the later-period applicability is not strong.
The antibacterial peptide A is a linear antibacterial peptide containing lysine or arginine and is BMAP-18, LL37 or magainin, wherein the antibacterial peptides BMAP-18, LL37 and magainin are all purchased from Kangtai Biotechnology (Beijing) Co., Ltd.
The number m of the antibacterial peptides is 2-32.
The invention also provides a preparation method of the antibacterial polypeptide, which comprises the following steps:
s100, reacting polyamide-amine type dendritic macromolecules with macromolecules at the tail ends thereof as amino groups with glutaraldehyde to obtain glutaraldehyde-activated polyamide-amine type tree molecules;
s200, removing redundant glutaraldehyde through ultrafiltration, then reacting the polyamide-amine type tree molecules with antibacterial peptide for a certain time, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
In a preferred embodiment, the reaction time of step S100 is 2-12 hours, and the temperature is 10-40 ℃; the reaction of step S200 is carried out for 2-8 hours at a temperature of 10-40 ℃.
The invention also provides another preparation method of the antibacterial polypeptide, which comprises the following steps:
s1000, reacting the polyamide-amine dendrimer of which the tail end is carboxyl and carboxylate with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide to obtain active ester of the polyamide-amine dendrimer;
s2000, removing redundant 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide through ultrafiltration, reacting the active ester with the antibacterial peptide for a certain time, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
In a preferred embodiment, the reaction time of step 1 is 2-5h, and the temperature is 10-40 ℃; the reaction of the step 2 is carried out for 2 to 12 hours at the temperature of between 10 and 40 ℃,
wherein, if the reaction time is less than 2 hours, the glutaraldehyde-activated dendrimer does not completely react with the antimicrobial peptide. The temperature is lower than 10 ℃, and the reaction efficiency is too low; the reaction time is more than 8 hours, and the temperature is higher than 40 ℃ to influence the bacteriostatic activity of the antibacterial peptide.
Finally, the invention provides an application of the poly antibacterial peptide in preparing livestock and poultry feed additives or immunopotentiators.
The following describes 2 specific embodiments of the present invention with reference to the drawings of the specification.
Example 1
The preparation method of the dendritic antibacterial peptide comprises the following steps:
s1000, reacting the 1.5 th generation carboxyl-terminal polyamide-amine type dendritic macromolecule with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) in 50mM morpholinoethanesulfonic acid buffer solution (pH 6.0) for 2 hours to obtain active ester of the polyamide-amine type dendritic macromolecule;
s2000, removing redundant 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide through ultrafiltration, then adding antibacterial peptide BMAP-18, reacting for 5 hours, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
As shown in FIG. 2, gel electrophoresis of the peptides and peptides shows, the peptides have been successfully linked to dendrimers, each dendrimer having 3-5 BMAP-18 peptides linked thereto, so that the number of active sites of the prepared peptides is increased by 3-5 times as compared with the original peptides.
Spotting experiment:
three test strains (gram positive bacteria, staphylococcus aureus 26001; gram negative bacteria, escherichia coli C600 and salmonella 13076) were applied to LB medium plates, and then diluted stepwise to prepare 10mg/mL, 5mg/mL, 2.5mg/mL, 1.25mg/mL, 0.625mg/mL and 0.3125mg/mL of the antimicrobial peptide and 1mg/mL, 0.5mg/mL, 0.25mg/mL, 0.125mg/mL, 0.0625mg/mL and 0.03125mg/mL of the antimicrobial peptide, respectively, and the spotting results are shown in fig. 3(a) to fig. 5 (b). The minimum concentration of antiplaque peptide was 0.625mg/mL and the minimum concentration of antiplaque peptide was 0.0625mg/mL, 10-fold lower than that of the antibacterial peptide, across the three strains tested.
The dendritic polymeric antibacterial peptide liquid preparation prepared in the embodiment 1 and the powder produced by spray drying, freeze drying and the like can be prepared into livestock and poultry feed additives or livestock and poultry disease prevention and treatment preparations.
60 healthy 1-day-old male white-feather chickens with similar body weights are selected and randomly divided into three treatment groups, wherein each group comprises 20 chickens, an experiment group I feeds a feed added with one thousandth of antibacterial peptide powder, an experiment group II feeds a feed added with one thousandth of antibacterial peptide powder, and an experiment group III feeds a basic feed. On day 35, chicken sera were taken and tested for chicken immune markers interferon (γ -IFN), chicken immunoglobulin A (SIGA) and chicken interleukin-2 (IL-2) using an ELISA kit, as detailed in Table 1. The result shows that the serum three immune indexes of gamma-IFN, SIgA and IL-2 of the experimental group added with the antibacterial peptide and the antibacterial peptide are all obviously higher than that of the experimental group fed with the basic feed, wherein the immune index fed with the antibacterial peptide is the highest, which shows that the antibacterial peptide can be used as a feed additive for livestock and poultry, and can enhance the non-specific immunity of animals and prevent diseases.
TABLE 1 immunization index of the chicks of each group
Example 2
The preparation method of the dendritic antibacterial peptide comprises the following steps:
s100, reacting the 2 nd generation polyamide-amine type dendrimer with the tail end being an amino terminal with a 5% glutaraldehyde solution for 2 hours to obtain a glutaraldehyde-activated polyamide-amine type dendrimer;
s200, removing redundant glutaraldehyde through ultrafiltration, adding antibacterial peptide BMAP-18 into the glutaraldehyde-activated polyamide-amine type dendritic macromolecules to react for 2 hours, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
Then, the antibacterial peptide and the antibacterial peptide prepared in example 2 were subjected to spotting test, and the minimum inhibitory concentration is shown in table 2 below:
TABLE 2 comparison of the dendritic polymeric antibacterial peptides prepared in example 2 with the antibacterial peptide spotting experiments
As can be seen from the above table, the antibacterial peptides are shown in the following formulas of Staphylococcus aureus M26001, Escherichia coli C600, Salmonella 13076, Salmonella pullorum S96116, and Escherichia coli O157: the minimum inhibitory concentration of H7 is 0.3125mg/mL, while the minimum inhibitory concentration of the poly antibacterial peptide prepared in example 2 against the above five bacteria is 0.03125mg/mL, which is 10 times lower than that of the antibacterial peptide.
The dendritic polymeric antibacterial peptide liquid preparation obtained in the above example 2 and the powder produced by spray drying, freeze drying and the like can be made into livestock and poultry feed additives or livestock and poultry disease prevention and treatment preparations.
Example 3 dendritic polymeric antibacterial peptides were prepared by the following steps:
s1000, reacting the 2.5 th generation carboxyl-terminal polyamide-amine type dendritic macromolecule with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) in 50mM morpholinoethanesulfonic acid buffer solution (pH 6.0) for 5 hours to obtain active ester of the polyamide-amine type dendritic macromolecule;
s2000, removing redundant 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide through ultrafiltration, adding the antibacterial peptide LL37, reacting for 10 hours, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide. Then, the antibacterial peptide and the prepared poly antibacterial peptide are subjected to a sample application experiment, and the minimum inhibitory concentration is shown in the following table 3:
the dendritic polymeric antibacterial peptide liquid preparation obtained in the above example 3 and the powder produced by spray drying, freeze drying and the like can be made into livestock and poultry feed additives or livestock and poultry disease prevention and treatment preparations.
TABLE 3 comparison of the dendritic polymeric antibacterial peptides prepared in example 3 with the antibacterial peptide spotting experiments
Example 4
The preparation method of the dendritic antibacterial peptide comprises the following steps:
s100, reacting the 3 rd generation polyamide-amine type dendrimer with the tail end being an amino terminal with a 5% glutaraldehyde solution for 2 hours to obtain a glutaraldehyde-activated polyamide-amine type dendrimer;
s200, removing redundant glutaraldehyde through ultrafiltration, adding antibacterial peptide magainin into the glutaraldehyde-activated polyamide-amine type dendritic macromolecules for reaction for 6 hours, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
Then, the antibacterial peptide and the prepared poly antibacterial peptide are subjected to a sample application experiment, and the minimum inhibitory concentration is shown in the following table 4:
table 4 comparison of the dendritic polymeric antibacterial peptides prepared in example 4 with the antibacterial peptide spotting experiments
The dendritic polymeric antibacterial peptide liquid preparation obtained in the above example 4 and the powder produced by spray drying, freeze drying and the like can be made into livestock feed additives or livestock disease prevention and treatment preparations.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Sequence listing
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Claims (10)
1. A dendritic polyacidin of the formula: DAmWherein D represents a polyamidoamine-type dendrimer, A represents an antibacterial peptide, and m represents the number of the antibacterial peptides.
2. The peptide according to claim 1, wherein the polyamidoamine dendrimer terminal group is preferably an amino group, a carboxyl group, or a carboxylate.
3. The antibacterial polypeptide of claim 1, wherein the polyamidoamine dendrimer has 1-3 generations.
4. The poly antibacterial peptide according to claim 1, wherein the antibacterial peptide a is a linear antibacterial peptide containing lysine or arginine.
5. The peptide of claim 1, wherein the number m of said peptides is any integer value from 2 to 32.
6. A process for the preparation of a poly-antibacterial peptide as claimed in any one of claims 1 to 5, which process comprises the steps of:
s100, reacting polyamide-amine type dendritic macromolecules with macromolecules at the tail ends thereof as amino groups with glutaraldehyde to obtain glutaraldehyde-activated polyamide-amine type tree molecules;
s200, removing redundant glutaraldehyde through ultrafiltration, then reacting the polyamide-amine type tree molecules with antibacterial peptide for a certain time, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
7. The method according to claim 6, wherein the reaction time of step S100 is 2 to 12 hours and the temperature is 10 to 40 ℃; the reaction of step S200 is carried out for 2-8 hours at a temperature of 10-40 ℃.
8. A process for the preparation of a poly-antibacterial peptide as claimed in any one of claims 1 to 5, which process comprises the steps of:
s1000, reacting the polyamide-amine dendrimer of which the tail end is carboxyl and carboxylate with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide to obtain active ester of the polyamide-amine dendrimer;
s2000, removing redundant 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide through ultrafiltration, reacting the active ester with the antibacterial peptide for a certain time, and removing redundant antibacterial peptide through ultrafiltration to obtain the dendritic polymeric antibacterial peptide.
9. The method for preparing a poly (antibacterial peptide) according to claim 8, wherein the reaction time of step 1 is 2-5 hours, and the temperature is 10-40 ℃; the reaction of step 2 is carried out for 2-12 hours at a temperature of 10-40 ℃.
10. Use of the poly antibacterial peptide according to any one of claims 1 to 5 in the preparation of a feed additive or an immunopotentiator for livestock and poultry.
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