Pseudomonas rhodesiae PYQ4 for highly producing exopolysaccharide and preparation method and application of exopolysaccharide

A technology of Pseudomonas Roche and extracellular polysaccharide is applied in the field of microorganisms to achieve the effects of repairing post-sun damage, low price, and simple and easy-to-obtain active ingredients

Active Publication Date: 2019-03-19
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, research on the application of microbial exopolysaccharides in improving skin properties has increased, such as the use of phosphorylated polysaccharides produced by Lactococcus lactis as moisturizers and whitening agents (Japanese Patent Application Publication No. 9-249524 , Japanese Patent Laying-Open No. 10-251140), a method of using polysaccharide components produced by Lactobacillus strains as an anti-inflammatory agent (Japanese Patent Laid-Open No. 7-70209 communique), and the exopolysaccharide isolated from lactic acid bacteria fermented milk prevents hairless Ultraviolet-induced skin damage in mice (International Journal of Molecular Sciences, 2017), etc., but there is no report on the use of Pseudomonas exopolysaccharide as an after-sun repair ingredient

Method used

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  • Pseudomonas rhodesiae PYQ4 for highly producing exopolysaccharide and preparation method and application of exopolysaccharide
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  • Pseudomonas rhodesiae PYQ4 for highly producing exopolysaccharide and preparation method and application of exopolysaccharide

Examples

Experimental program
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example 1

[0019] Example 1: Extraction and Content Determination of Exopolysaccharides Produced by Pseudomonas Roche PYQ4

[0020] An appropriate amount of the above strains were picked from the sugar-producing solid medium and inoculated into the sugar-producing fermentation medium. The fermentation broth was obtained by shaking culture for 48h in a shaker with a rotational speed of 150rpm at 30°C. figure 1 It is the plate culture character of the strain PYQ4 of the present invention.

[0021] Among them, the used sugar-producing solid medium components are: sucrose 20g / L, tryptone 10g / L, yeast powder 5g / L, Na 2 HPO 4 ·12H 2 O 3g / L, agar 20g / L, the balance is distilled water. The components of the sugar-producing fermentation broth used are: sucrose 50g / L, tryptone 5g / L, yeast powder 1g / L, Na 2 HPO 4 ·12H 2 O 3g / L, the balance is distilled water.

[0022] The fermentation broth was centrifuged at 4000 g for 10 min to remove bacterial cells to obtain a supernatant. 1 / 4 volume o...

example 2

[0024] Example 2: Identification of the microbial strains of the present invention

[0025] This strain is red by Gram staining and is a Gram-negative bacteria. The total DNA of the strain of the present invention was extracted according to the steps of the Takara Bacteria Genomic DNA Extraction Kit, and the 16S rDNA gene of the strain was amplified with the universal primers 27F and 1492R. Sequencing, and bacterial species identification by determining the sequence of 16S rDNA. The obtained sequence results were compared by Blast at NCBI, and the recognized standard sequence data homologous to the 16S rDNA of the strain was obtained from the database of GenBank. The MEGA software was used to calculate the sequence similarity and the Neighbor-Joining algorithm was used as the system. Developmental Analysis.

[0026] The constructed phylogenetic tree of the strain is as follows figure 2 As shown, it has the highest homology with Pseudomonas rhodesiae, so the strain describe...

example 3

[0027] Example 3: Application of above-mentioned polysaccharide as post-sun damage repairing agent

[0028] HaCaT cells in log phase were collected and seeded in a 96-well plate, 5000 cells per well, at 37°C, 5% CO. 2 Cultivated in an incubator. The cells were divided into control group, UV model group, polysaccharide group (1mg / mL), ethylhexyl methoxycinnamate (MCX) group (8%) and ascorbic acid (Vc) group (1mg / mL), of which the latter two. The group was the positive control, and each group was replicated with 3 wells. After the cells were cultured for 24 h, the medium was changed to phosphate buffered saline (PBS) and then irradiated with UV, and the control group was wrapped with tin foil to protect from light. Aspirate PBS, add the medium, the medium of the polysaccharide group contains 1 mg / mL of the above-mentioned strains to produce extracellular polysaccharides, the medium of the MCX group contains 8% ethylhexyl methoxycinnamate, and the Vc group is cultured The base...

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Abstract

The invention discloses Pseudomonas rhodesiae PYQ4 for highly producing exopolysaccharide and a preparation method and application of exopolysaccharide. The strain is classified and named as Pseudomonas rhodesiae PYQ4 and is preserved in China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.16674. The preparation method comprises the following steps: isolation culture and 16S rDNA authentication of the strain, the acquisition of a strain fermentation product, namely exopolysaccharide and application of the strain fermentation product to the repairingof sun damage. By virtue of the exopolysaccharide produced by virtue of the Pseudomonas rhodesiae disclosed by the invention, the proliferation activity of sun-damaged cells can be remarkably improved, and the exopolysaccharide has an obvious effect to the repairing of sun damage, has the characteristics of low cost, simplicity and easiness in preparation and the like when being taken as a naturalproduct of microbial fermentation, can be applied to the preparation of relevant sunning repairing agents and has good market application prospects.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a Pseudomonas Roche strain PYQ4 with high production of extracellular polysaccharide, and the use of the produced extracellular polysaccharide in post-sun repair. Background technique [0002] With the increase of air pollution year by year, the ozone layer has been severely damaged, and the ultraviolet radiation to the earth's surface has increased significantly, affecting people's quality of life. At present, the damage effect of UV on organisms and its protection have become a hot issue of global concern. [0003] The skin is the body's first line of defense and biological barrier against various external physical and chemical stimuli, among which UV is one of the important harmful factors of skin contact. Studies have shown that overexposure to UV radiation is the main cause of skin photodamage, which can cause erythema, inflammation, pigmentation, and mor...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/04A61K8/73A61Q19/00C12R1/38
CPCA61K8/73A61K2800/85A61Q19/004C12P19/04C12N1/205C12R2001/38
Inventor 周文文汪旖清
Owner ZHEJIANG UNIV
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