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Application of Plectasiacin in Inhibiting Streptococcus agalactiae

A technology of Plectasin and Streptococcus lactis, applied in antibacterial drugs, medical preparations containing active ingredients, peptide/protein components, etc., can solve unknown problems

Active Publication Date: 2017-01-18
GUANGDONG HINAPHARM PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although the existing information discloses that Plectasin can inhibit the growth of various bacteria, whether it can inhibit the growth of Streptococcus agalactiae and whether it can be used for the treatment of mastitis is still unknown

Method used

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  • Application of Plectasiacin in Inhibiting Streptococcus agalactiae
  • Application of Plectasiacin in Inhibiting Streptococcus agalactiae
  • Application of Plectasiacin in Inhibiting Streptococcus agalactiae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation method of embodiment 1 plectasin protein

[0018] In this example, a method of exogenously expressing the target protein plectasin using Lactococcus lactis NZ9700 as the host was used to prepare high-purity plectasiacin.

[0019] 1. Construction of recombinant strains

[0020] The nucleotide sequence of the Usp45 protein signal peptide was cloned from Lactococcus lactis MG1363, and the Usp45 gene fragment was connected with the shuttle plasmid pMG36e to form plasmid pGMN1; the plasmid pGMN1 was connected with the Pnis-MCS segment gene amplified from the lactic acid bacteria plasmid pNZ8408, Construction of plasmid pGMN2; artificially synthesized lactic acid bacteria codon-biased plectasin gene fragment, the optimized plectasin base sequence is: GGT TTT GGT TGT AAT GGT CCA TGG GAT GAA GAT GATATG CAA TGT CAT AAT CAT TGT AAA TCA ATT AAA GGT TAT AAA GGT GGT TAT TGT GCTAAA GGT GGT TTT GTT TGT AAA TGT TAT (SEQ ID NO: 1), the optimized Plectasin gene fragment ...

Embodiment 2

[0033] Example 2 Inhibitory effect of plectasin protein on streptococcus agalactiae

[0034]Take the above-mentioned mycelia protein purified by high-performance liquid chromatography, make it into a concentration of 1 μg / μL, and place the sample on the agar plate containing Streptococcus agalactiae; at the same time, the plate also contains ampicillin Amp and sterile water. Serve as positive and negative controls respectively; culture overnight at 37°C and observe the results.

[0035] The result is as image 3 As shown, there were obvious inhibition zones in all points where there were different volumes of mycelia solution and ampicillin, and the places where there was sterile water also listed the inhibition circles, indicating that mycelia had the ability to inhibit Streptococcus agalactiae effect.

Embodiment 3

[0036] Example 3 The minimum inhibitory concentration MIC test of Plectasia to Streptococcus agalactiae

[0037] The Streptococcus agalactiae liquid is adjusted with physiological saline to detect at a wavelength of 625nm, and the absorption value is 0.09-0.10, then diluted with sterilized broth in different proportions, and shaken evenly. Add different amounts of the above-mentioned purified plectasin or Amp into each test tube, so that the concentration of plectasin in each sample test tube is 1000 μg / mL, 100 μg / mL, 10 μg / mL, 1 μg / mL, 0.5μg / mL; the concentration of Amp in each sample tube is 500μg / ml, 250μg / ml, 125μg / ml, 62μg / ml, 31μg / ml, 16μg / ml, 8μg / ml, 4μg / ml, 2μg / ml ml, 1 μg / ml, 0.5 μg / ml, 0.25 μg / ml, 0.125 μg / ml, 0.062 μg / ml, 0.031 μg / ml. Then place it under the condition of 37° C. and incubate for 12 hours.

[0038] The test results showed that the MIC of Plectasia to Streptococcus agalactiae was 1 μg / mL of the purified Plectasia protein, and the MIC of Amp against S...

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Abstract

The invention discloses an application of plectasin in inhibiting streptococcus agalactiae. The plectasin is found to be good at inhibiting the streptococcus agalactiae through bacteriostasis experiments; the plectasin is further prepared into corresponding agentia, such as plectasin gel and plectasin emulsion, the agentia achieve good curative effects for mastitis, wherein the antibacterial anti-inflammatory effect of the gel is better, the action time lasts longer, the moisturizing effect is better, action sites can be effectively isolated from making contact with air, and a wound is prevented from infection and inflammation. The plectasin can be used for preparing bacteriostat of the streptococcus agalactiae and further used for curing the mastitis, and especially for the mastitis caused by the streptococcus agalactiae which is difficult to cure at present, and a good curative effect is achieved.

Description

technical field [0001] The invention relates to the application of plectasin in inhibiting Streptococcus agalactiae. Background technique [0002] β-hemolytic streptococci are divided into 18 groups from A to T, among which, agalactiae is divided into group B, which is the most common pathogenic streptococcus except group A hemolytic streptococci. Streptococcus agalactiae (S. agalactiae) is the pathogen of mastitis, often present in the skin, teats and udders of dairy cows, through towels, water, milkers' hands, milking cups, etc. used to scrub udders. Infectious pathogenic bacteria are more harmful. Treatment during milking period can control Streptococcus agalactiae, but it cannot immediately solve the problem of excessive somatic cell count in milk during lactation period, which will affect the sales of milk. The spread of dairy cow mastitis is very fast, and it has been a major problem that has plagued the development of the dairy industry. The incidence of bovine mast...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61P31/04A61P15/14
Inventor 周玉岩孙丹丹逯佩凤李洪金
Owner GUANGDONG HINAPHARM PHARMA CO LTD
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