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Artificial synthesis method and application of antibacterial and insect-resistant peptide

A synthesis method and reaction technology are applied in the field of artificial synthesis of antibacterial and insecticidal peptides, which can solve problems such as insufficient control methods for resistant pests, and achieve the effects of efficiently inhibiting growth and development, solving control problems, and broad-spectrum bactericidal properties.

Active Publication Date: 2021-07-23
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to overcome the deficiencies of the existing methods of drug-resistant strain-resistant pest control, and to provide an artificial synthesis method of antibacterial and insect-resistant peptides and its application

Method used

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  • Artificial synthesis method and application of antibacterial and insect-resistant peptide
  • Artificial synthesis method and application of antibacterial and insect-resistant peptide
  • Artificial synthesis method and application of antibacterial and insect-resistant peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. D-type peptide thermal polymerization method:

[0038] (1) Formulate the corresponding amino acids into a 3g system according to the formula ratio (see Table 1), and set up three parallels for each formula. Add 4.8ml of peanut oil and 0.6ml of phosphoric acid to every 3g of amino acid system, mix well, place in an oven at 150°C, and thermally polymerize for 4 hours.

[0039] (2) Wash and centrifuge three times with absolute ethanol, the centrifugal speed is 6000rpm, 5min;

[0040] (3) Dry in an oven at 60°C for 2 hours.

[0041] The D-type peptide formula of table 1 embodiment 1

[0042] serial number Recipe(w:w) 107 D-Glu:D-Lys hydrochloride=2:1 109 D-Glu:L-Pro=2:1 112 D-Lys hydrochloride:D-Asp:L-Thr=1:1:8

[0043] 2. Inhibition zone test of D-peptide

[0044] S1. Preparatory work such as preparation of culture medium, sterilization of culture medium and petri dish.

[0045] S2. Culture Staphylococcus aureus Y5, and measure col...

Embodiment 2

[0063] The MIC determination of D-type peptides to different bacteria under the low-salt low-protein concentration medium condition of embodiment 2

[0064] 1. Experimental materials

[0065] Peptides: 107, 109, 112 in Example 1, specifically 107-1, 107-2, 107-3, 109-1, 109-2, 109-3, 112-1, 112-2, 112-3;

[0066] Concentration: 0.25, 0.125, 0.0625mg / ml.

[0067] Strains: Ralstonia solanacearum, Pseudomonas aeruginosa ATCC.

[0068] LSLP medium formula: sodium chloride, 0.0001%; ​​yeast powder, 0.05%; peptone, 0.1%.

[0069] 2. Pre-experimental preparation

[0070] (1) Activated bacteria: first activate the preserved bacteria, and after activation, draw a line, take a single colony and add it to 5ml LB liquid medium for shaking culture until the OD value reaches about 0.6 (37°C, 220 rpm), then connect it to solid LB Plates were counted in the medium, and the bacterial solution was stored at 4°C for MIC experiments.

[0071] (2) Dilute the bacterial solution to 10 with ster...

Embodiment 3D

[0085] The effect of embodiment 3D type peptide on beet armyworm

[0086] 1. Experimental materials: beet armyworm, disposable lunch box, gauze, tweezers, insect feed.

[0087] 2. Rearing of beet armyworm

[0088] (1) The larva rearing condition is 27°C, dark: light (D:L) = 14:10;

[0089] (2) Disposable lunch boxes or petri dishes for feeding larvae need to be irradiated for at least 30 minutes under ultra-clean bench ultraviolet light;

[0090] (3) Use tweezers (disinfected) to carefully place insect larvae less than 3 instar (body length < 1cm) in a petri dish for every 25 larvae;

[0091] (4) After roasting and disinfecting the feed on the alcohol lamp with a cutting knife, cut the feed into thin slices about 2mm thick. Just place 3 to 4 pieces of feed in the petri dish;

[0092] (5) Observe the growth of larvae every day and add feed, pay attention to clean the feces or replace with new petri dishes.

[0093] 3. Effect experiment of D-type peptide on beet armyworm

...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to an artificial synthesis method and application of antibacterial and insect-resistant peptide. The synthesis method of D-type antibacterial and insect-resistant peptide comprises the following steps: S1, taking a proper amount of amino acid raw materials, adding peanut oil and phosphoric acid, and carrying out thermal polymerization reaction, wherein the volume ratio of the peanut oil to the phosphoric acid is (0-8):(0-1); and S2, taking out the polymer, washing the polymer with absolute ethyl alcohol, and carrying out centrifuging and drying to obtain the D-type antibacterial and insect-resistant peptide, wherein the amino acid raw materials are D-glutamic acid and D-lysine hydrochloride, or D-glutamic acid and L-proline, and the weight ratio of the D-glutamic acid to the D-lysine hydrochloride or the weight ratio of the D-glutamic acid to the L-proline is 2:1. The D-type antibacterial and insect-resistant peptide prepared by the synthesis method has the characteristics of high efficiency and broad-spectrum sterilization, can efficiently inhibit growth and development of beet armyworm, and has the characteristic of high-efficiency insect resistance. The antibacterial and insect-resistant peptide prepared by the method has extremely high industrial value and application potential in the aspects of inhibiting drug-resistant bacteria and drug-resistant pests, preparing novel antibiotics and novel insect-resistant drugs and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an artificial synthesis method of an antibacterial and insect-resistant peptide and its application. Background technique [0002] In recent years, the long-term abuse of antibiotics has led to more and more inflammatory diseases caused by drug-resistant strains, which has seriously affected global public health security, and the number of new antibiotics that have successfully entered clinical applications has been gradually shrinking, making it difficult to find alternatives to antibiotics. become a priority for people. As one of the oldest defense mechanisms of organisms, antimicrobial peptides have very important physiological functions. They can effectively kill bacteria, fungi, viruses and protozoa, and become an effective protective barrier to protect the host from pathogens. Studies have shown that antimicrobial peptides are effective against a variety of drug-re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K2/00A01N37/46A01N43/36A01P7/04A01P1/00
CPCC07K2/00A01N37/46A01N43/36Y02A50/30
Inventor 刘秋云何建国翁少萍杨凯贺竹梅高嘉阳周艳超戴丹玲潘梦佳
Owner SUN YAT SEN UNIV