CN115536738A - Application of cobra antibacterial peptide OH-CATH30 in treating enteritis of fishes - Google Patents
Application of cobra antibacterial peptide OH-CATH30 in treating enteritis of fishes Download PDFInfo
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- CN115536738A CN115536738A CN202210381228.4A CN202210381228A CN115536738A CN 115536738 A CN115536738 A CN 115536738A CN 202210381228 A CN202210381228 A CN 202210381228A CN 115536738 A CN115536738 A CN 115536738A
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Abstract
The invention relates to application of cobra antibacterial peptide OH-CATH30 in treating fish enteritis, belonging to the technical field of biological medicines. The molecular weight of the cobra antibacterial peptide OH-CATH30 is 4693.71Da, the isoelectric point is 12.08, and the amino acid sequence is KRFKKFFKKLKNSVKKRAKKFFKKPRVIGVSHIPHHH; application of antibacterial peptide OH-CATH30 of Ophiophagus hannah in treating fish enteritis is provided. The invention has the following effects: the cobra antibacterial peptide OH-CATH30 is mixed and stirred with fish feed, and the cod with enteritis is fed to effectively cure the enteritis of the cod.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to application of cobra antibacterial peptide OH-CATH30 in treating fish enteritis.
Background
The fish enteritis is bacterial enteritis caused by intestinal type punctate aeromonas infection, is the most common fish disease, can be attacked from fish species to adult fishes, and is particularly easy to attack by eating fishes such as grass carp, crucian and the like. The disease begins to spread at the water temperature of more than 18 ℃, is in a flow peak period at the temperature of 25-30 ℃, is frequently complicated with bacterial gill rot disease, red skin disease and the like, and is one of the more serious diseases in cultured fishes.
The use of terramycin, which is a broad spectrum antibiotic, is essential for the treatment of fish enteritis, however, the use of antibiotics has greatly contributed to the increase of antibiotic-resistant pathogens, accompanied by the use of antibiotics, especially the abuse of antibiotics, which has led to the gradual increase of bacterial resistance to antibiotics.
The cobra antibacterial peptide OH-CATH30 consists of 34 amino acid sequences. The application of the antibacterial peptide OH-CATH30 of the king cobra in the process of treating fish enteritis is researched by taking the antibacterial peptide OH-CATH30 of the king cobra as a research object.
Disclosure of Invention
The invention also aims to provide the application of the king cobra antimicrobial peptide OH-CATH30 in the process of treating fish enteritis, which can effectively cure fish enteritis, regulate the immunity level of fish organisms and improve feed conversion efficiency.
The invention is realized by the following technical scheme: an antibacterial peptide OH-CATH30 of king cobra, wherein the antibacterial peptide OH-CATH30 of king cobra consists of 34 amino acid sequences; the molecular weight of the cobra antibacterial peptide is 4693.71Da, the isoelectric point is 12.08, and the amino acid sequence of the cobra antibacterial peptide OH-CATH30 is KRFKKFFKKNSVKKRAKKFFKKPRVIGVSSIPHTHHHH.
Further, the Ophiophagus fasciatus antimicrobial peptide OH-CATH30 is obtained by a pichia pastoris fermentation method.
The invention relates to application of an antibacterial peptide OH-CATH30 of king cobra in treating enteritis of fishes.
The invention has the advantages that: the antibacterial activity of the obtained antibacterial peptide OH-CATH30 gene engineering expression product of the king cobra is identified, and an experiment for treating enteritis is carried out, wherein the experiment shows that the antibacterial peptide OH-CATH30 of the king cobra is mixed and stirred with fish feed, and the cod with enteritis is fed to the cod, so that the enteritis of the cod can be effectively cured.
Drawings
FIG. 1 control group pictures of pollack;
FIG. 2 Experimental group pictures of pollack;
FIG. 3 control group pictures of intestinal epithelial tissue of pollack;
fig. 4 experimental group pictures of intestinal epithelial tissue of pollack.
Detailed Description
1. The antibacterial peptide OH-CATH30 of king cobra is from the venom gland of king cobra (Ophiophagus hannah).
2. Vectors and host cells: both the pPIC9K plasmid and pichia pastoris GS115 strain were purchased from the plasmid platform of\28156ling.
3. The bacterial strains are purchased from China general microbiological culture Collection center.
Unless otherwise indicated, the reagents used in the following examples of the present invention are all analytically pure reagents and are commercially available from conventional sources.
The present invention is further described below, and the scope of protection of the present invention is not limited to the following:
example 1: preparation of king cobra antibacterial peptide OH-CATH30
S01, constructing an expression vector pPIC9K-OH-CATH30 of the Elaphe carinata antimicrobial peptide OH-CATH30, and specifically operating as follows: PCR amplifying an antibacterial peptide OH-CATH30 gene of the king cobra to obtain a target gene fragment of the antibacterial peptide OH-CATH30, and connecting the OH-CATH30 gene fragment with a pPIC9K vector to obtain a pPIC9K-OH-CATH30 recombinant vector;
s02, transforming the recombinant vector obtained in the step S01 into host cell Pichia pastoris GS115, and carrying out induction expression on the host cell to obtain an expression product;
s03, separating and purifying the expression product obtained in the step S02, wherein the specific operation is as follows: and (3) firstly centrifuging the expression product, collecting supernatant, dialyzing, and finally purifying by affinity chromatography to obtain the recombinant polypeptide OH-CATH30.
Example 2: preparation of king cobra antibacterial peptide OH-CATH30 as fish feed additive
(1) Selecting 30kg of feed of the 3# pollock in the market;
(2) Adding the cobra antibacterial peptide OH-CATH30 polypeptide sample obtained by the method in the embodiment 1 into feed according to the proportion of 0.5mg/kg, and uniformly mixing to obtain the feed containing the cobra antibacterial peptide OH-CATH30 polypeptide.
Example 3: experimental effect of using king cobra antibacterial peptide OH-CATH30 as fish feed additive for treating pollock enteritis
(1) Selecting 50 pollack fishes with the same weight, wherein the weight of each fish is 200g +/-10 g;
(2) And performing a toxicity attacking experiment on the pollack. The toxicity counteracting experiment adopts intraperitoneal injection method, and the injection is 1.0 multiplied by 10 8 0.1mL of enterobacter punctatus at cfu/mL concentration. Feeding at a daily bait feeding rate of 3% at a water temperature of 25 ℃, feeding for 3 times a day, and observing the cod with obviously reduced ingestion, namely the cod with enteritis.
(3) Setting the test group and the control group;
experimental groups: selecting 10 pollock suffering from enteritis, feeding the pollock of the experimental group with the uniformly mixed feed containing the king cobra antimicrobial peptide OH-CATH30 polypeptide in the example 2, reducing the daily feeding rate to 1% under the condition of 25 ℃ water temperature so as to ensure that the medicine bait can be completely ingested by the sick fish, feeding for 3 times a day, and continuously feeding for 10 days.
Control group: 10 pollock fishes suffering from enteritis are selected, the pollock fishes are fed by the same common fish feed without OH-CATH30, the feeding rate and the feeding frequency per day are the same as those of the experimental group, and the pollock fishes are continuously fed for 10 days.
(4) After 10 days, the cod of the experimental and control groups was dissected.
As shown in fig. 1, it was found that: the 10 silver cod fishes in the control group have swelling and flatulence in intestinal tracts, while the 10 silver cod fishes in the experimental group have normal intestinal tracts after being continuously fed with feed containing the king cobra antimicrobial peptide OH-CATH30 for 10 days, and the intestinal tracts are full of food and have no swelling and flatulence.
(5) And respectively slicing the burbot of the experimental group and the burbot of the control group in the intestinal tract, and observing the condition of the intestinal tract epithelial tissue. As shown in FIG. 2, the intestinal epithelial tissue of the silver cod in 10 control groups was disintegrated, while the intestinal epithelial tissue of the silver cod in the experimental group was intact.
The pollock feeding experiment shows that the cod fed with the cobra antibacterial peptide OH-CATH30 mixed material can effectively cure the enteritis of the pollock and restore the intestinal epithelial tissue structure of the pollock to be complete.
Sequence listing
<110> Jiangsu hypertonic Jun Biotechnology Co., ltd
<120> application of king cobra antibacterial peptide OH-CATH30 in treatment of enteritis of fishes
<130> 20220411
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 105
<212> DNA
<213> King cobra (Ophiophagus hannah)
<400> 1
aagagattca agaaattttt caagaagctg aagaacagcg tgaagaaacg tgccaagaaa 60
ttcttcaaga agccgagggt catcggagtc tccatcccct tctaa 105
<210> 2
<211> 40
<212> PRT
<213> Elaphe carinata (Ophiophagus hannah)
<400> 2
Lys Arg Phe Lys Lys Phe Phe Lys Lys Leu Lys Asn Ser Val Lys Lys
1 5 10 15
Arg Ala Lys Lys Phe Phe Lys Lys Pro Arg Val Ile Gly Val Ser Ile
20 25 30
Pro Phe His His His His His His
35 40
Claims (2)
1. The cobra antibacterial peptide OH-CATH30 is characterized in that the cobra antibacterial peptide OH-CATH30 is obtained by a pichia pastoris fermentation method, the molecular weight of the cobra antibacterial peptide OH-CATH30 is 4693.71Da, the isoelectric point of the cobra antibacterial peptide is 12.08, and the amino acid sequence of the cobra antibacterial peptide OH-CATH30 is KRFKKFFKFKFKFKFKFKSKVKKRAKKPRVIGVSHSHHH.
2. The use of the antibacterial peptide OH-CATH30 of the king cobra as claimed in claim 1, wherein the antibacterial peptide OH-CATH30 of the king cobra is used for treating enteritis in fish.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210381228.4A CN115536738A (en) | 2022-04-13 | 2022-04-13 | Application of cobra antibacterial peptide OH-CATH30 in treating enteritis of fishes |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210381228.4A CN115536738A (en) | 2022-04-13 | 2022-04-13 | Application of cobra antibacterial peptide OH-CATH30 in treating enteritis of fishes |
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| CN115536738A true CN115536738A (en) | 2022-12-30 |
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| CN202210381228.4A Pending CN115536738A (en) | 2022-04-13 | 2022-04-13 | Application of cobra antibacterial peptide OH-CATH30 in treating enteritis of fishes |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115536737A (en) * | 2022-04-13 | 2022-12-30 | 江苏亢钧生物科技有限公司 | Application of cobra antibacterial peptide OH-CATH30 in resisting aquatic animal pathogenic bacteria |
| CN116396372A (en) * | 2023-02-18 | 2023-07-07 | 青岛农业大学 | Peptide-loaded nanoparticle and application thereof in treatment of eye infection of pets |
-
2022
- 2022-04-13 CN CN202210381228.4A patent/CN115536738A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115536737A (en) * | 2022-04-13 | 2022-12-30 | 江苏亢钧生物科技有限公司 | Application of cobra antibacterial peptide OH-CATH30 in resisting aquatic animal pathogenic bacteria |
| CN115536737B (en) * | 2022-04-13 | 2023-10-03 | 江苏亢钧生物科技有限公司 | Application of cobra antibacterial peptide OH-CATH30 in preparation of bacterial growth inhibitor |
| CN116396372A (en) * | 2023-02-18 | 2023-07-07 | 青岛农业大学 | Peptide-loaded nanoparticle and application thereof in treatment of eye infection of pets |
| CN116396372B (en) * | 2023-02-18 | 2024-05-24 | 青岛农业大学 | Peptide-loaded nanoparticle and application thereof in treatment of eye infection of pets |
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