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Lipopeptid and derivative thereof, and preparation methods and application of lipopeptid and derivative of lipopeptid

A technology of derivatives and lipopeptides, applied in the field of lipopeptides and their derivatives that improve the antibacterial ability of the host, and its preparation, can solve problems such as the therapeutic effect of antibiotics on infection

Active Publication Date: 2012-09-12
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are more and more drug-resistant strains of microorganisms, and many antibiotics no longer play a good role in the treatment of microbial infections.

Method used

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  • Lipopeptid and derivative thereof, and preparation methods and application of lipopeptid and derivative of lipopeptid
  • Lipopeptid and derivative thereof, and preparation methods and application of lipopeptid and derivative of lipopeptid
  • Lipopeptid and derivative thereof, and preparation methods and application of lipopeptid and derivative of lipopeptid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1: Separation and purification of lipopeptides.

[0101] See Smyth, Thomas et al. Isolation and Analysis of Lipopeptides and High Molecular Weight Biosurfactants, 2010. The strain used was Staphylococcus epidermidis 1457.

[0102] (1) Staphylococcus epidermidis fermentation

[0103] ①Take out the preserved Staphylococcus epidermidis 1457 at -80°C, streak on tryptic soy broth (TSB) solid medium, and keep at 37°C for 16 hours until a single colony grows.

[0104] ②Pick a single colony and inoculate it in 20ml TSB liquid medium for culture at 37°C, 220rpm, for 16h.

[0105] ③Transfer the Staphylococcus epidermidis bacteria liquid cultured overnight to 200ml TSB liquid medium at 1%, and ferment at 37°C, 220rpm, for 16h.

[0106] (2) Acid precipitation in fermentation broth

[0107] ① Centrifuge the Staphylococcus epidermidis fermentation broth. 10000rpm, 4°C, 20min.

[0108] ② Adjust the supernatant of the Staphylococcus epidermidis fermentation broth after ce...

Embodiment 2

[0119] Example 2: Identification of lipopeptides.

[0120] Take 10 μl of the lipopeptide separated in Example 1 and spot it on a thin-layer chromatography plate, and the spotting position is 1 cm away from the lower edge. Place the chromatography plate in a chromatography cylinder presaturated with developing medium. The spreading agent is n-butanol: acetic acid: water = 4:2:1. Chromatography was stopped after the developing agent ran to 1cm from the edge of the chromatographic plate. Let dry in a fume hood. The board was then placed in water for 5 min, and then dried naturally in a fume hood. Observe the position of the lipopeptide on the plate. After completely drying, spray 0.25% ninhydrin on the plate, place it in an oven at 60°C, and observe the lipopeptide on the plate after 10 minutes. Experimental results such as figure 1 As shown, the lipopeptide forms an oil-expelling ring in the water phase, and after thin-layer chromatography, it is developed with water to pr...

Embodiment 3

[0121] Embodiment 3: lipopeptide HPLC analysis and molecular weight identification, the identification of polypeptide sequence

[0122] The lipopeptide separated in Example 1 was loaded onto a high-pressure liquid chromatography analyzer (provided by Shanghai Zhongke New Life Co., Ltd.). The mobile phase was gradient eluted with 0.1% TFA in water and 0.1% TFA in acetonitrile for 80 minutes, and then the eluted peaks were analyzed for molecular weight by peptide mass spectrometry. Analysis by HPLC, such as figure 2 As shown, three main peaks are obtained, and the peak time is 10min, 69min, and 70min respectively. After peptide mass spectrometry analysis, such as image 3 As shown, the peak time of 10min shows that there are two kinds of substances, and their molecular weights are 1437 and 1621 respectively. Such as Figure 4 As shown, the peak with a peak time of 69 min shows that there is a substance with a molecular weight of 2491. Such as Figure 5 As shown, the molec...

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Abstract

The invention provides lipopeptid which comprises a peptide chain and an aliphatic chain which are connected through peptide bonds, wherein the molecular weight of the lipopeptid is 2848Da, and the lipopeptid is linear, and can obviously introduce the expression of phylaxin, and effectively inhibit staphylococcus aureus infection so as to prevent or reduce skin infection. The invention also discloses a derivative of the lipopeptid and preparation methods and application of the lipopeptid and the derivative of the lipopeptid.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a lipopeptide and its derivatives for improving the antibacterial ability of a host, and a preparation method and application thereof. Background technique [0002] Since the advent of penicillin in the 1930s and 1940s, people have discovered many antibiotics such as cephalosporins, aminoglycosides, macrolides, and tetracyclines. Antibiotics play an important role in the treatment of microbial infections in humans. However, there are more and more drug-resistant strains of microorganisms, and many antibiotics no longer have a good therapeutic effect on microbial infections. Therefore, finding alternatives to antibiotics has become an increasingly urgent task for human beings. [0003] The lipopeptide is composed of two parts: a hydrophilic peptide chain and a lipophilic fatty acid chain, that is, about 10 polypeptides and fatty acid chains form a circular or linear lipop...

Claims

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Application Information

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IPC IPC(8): C07K14/31C12P21/02A61K38/16A61K8/64A61P31/04A61Q19/00A61Q19/10C12R1/45
CPCA61K8/64A61K38/00A61K47/542A61P31/04A61Q17/04C07K5/101C07K14/31
Inventor 李冬青雷虎李红泉赖玉平
Owner EAST CHINA NORMAL UNIV
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