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A high-yielding exopolysaccharide Pseudomonas roseri pyq4 and its polysaccharide preparation method and application

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

Active Publication Date: 2020-05-29
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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  • A high-yielding exopolysaccharide Pseudomonas roseri pyq4 and its polysaccharide preparation method and application
  • A high-yielding exopolysaccharide Pseudomonas roseri pyq4 and its polysaccharide preparation method and application
  • A high-yielding exopolysaccharide Pseudomonas roseri pyq4 and its polysaccharide preparation method and application

Examples

Experimental program
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example 1

[0019] Example 1: Extraction and content determination of exopolysaccharide produced by Pseudomonas roseri PYQ4

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

[0021] Among them, the components of the sugar-producing solid medium used are: sucrose 20g / L, tryptone 10g / L, yeast powder 5g / L, Na 2 HPO 4 12H 2 O 3g / L, agar 20g / L, and 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 above fermentation broth was centrifuged at 4000 g for 10 min to remove the bacteria to obtain the supernatant. Add 1...

example 2

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

[0025] This strain is red by Gram stain and is Gram-negative. According to the steps of TakaraBacteria Genomic DNA Extraction Kit (TakaraBacteria Genomic DNA Extraction Kit) to extract the total DNA of the strains described in the present invention, amplify the 16S rDNA gene of the bacteria with universal primers 27F and 1492R, recover the amplified products and carry out Sequencing, identification of bacterial species 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 GenBank database. The sequence similarity was calculated using MEGA software and the Neighbor-Joining algorithm (Neighbor-Joining) was used as the system developmental analysis.

[0026] The phylogenetic tree of the constructed strain is as follows: figure 2 As shown, the ho...

example 3

[0027] Example 3: Use of the above-mentioned polysaccharides as a repairing agent for sun damage

[0028] HaCaT cells in the logarithmic phase were collected and seeded in a 96-well plate, with 5000 cells per well, placed at 37°C, 5% CO 2 cultured in an incubator. 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), the latter two One group is the positive control, and each group has 3 wells repeated. After the cells were cultured for 24 hours, the culture medium was replaced with phosphate buffered saline (PBS) and then irradiated with UV light. The control group was wrapped in tinfoil to avoid light. Aspirate the PBS and add the culture medium, wherein the culture medium of the polysaccharide group contains 1 mg / mL of the prolific exopolysaccharides of the above strains, the culture medium of the MCX group contains 8% ethylhexyl methoxycinnamate, and the cul...

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Abstract

The invention discloses a high-yield extracellular polysaccharide Pseudomonas roseri PYQ4 and a preparation method and application of the polysaccharide. The strain is classified as Pseudomonas rhodesiae PYQ4, and is preserved in China General Microorganism Culture Collection and Management Center with the preservation number CGMCC No.16674. The invention includes the isolation and cultivation of the bacterial strain and the identification of 16S rDNA; the acquisition of the extracellular polysaccharide of the fermentation product of the bacterial strain and the application of the extracellular polysaccharide produced by the bacterial strain in repairing sun damage. The polysaccharide produced by Pseudomonas rosenbergii PYQ4 disclosed by the present invention can significantly improve the value-added activity of sun-damaged cells, has obvious sun-damage repairing effect, and as a natural product of microbial fermentation, it has the characteristics of low cost, simple and easy to obtain, etc. It can be applied to the preparation of related after-sun restoration agents, and has good market application prospects.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a high-yielding exopolysaccharide Pseudomonas roseri strain PYQ4 and the use of the produced exopolysaccharide in sun repair. Background technique [0002] With the increase of air pollution year by year, the ozone layer has been severely damaged, and the ultraviolet rays radiated to the earth's surface have increased significantly, affecting people's quality of life. At present, the harmful effect of UV on organisms and its protection have become a hot issue of global concern. [0003] Skin is the first line of defense and biological barrier for the human body to resist various physical and chemical stimuli from the outside world, among which UV is one of the important harmful factors of skin contact. Studies have shown that overexposure to ultraviolet radiation is the main cause of skin photodamage, which can cause erythema, inflammation, pigmentation, etc. UV ra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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