Extracellular polysaccharide high-yielding lactic acid bacterium BL21, and preparation method and application thereof

A technology of exopolysaccharides and lactic acid bacteria, which is applied in the field of microorganisms to achieve excellent safety effects

Active Publication Date: 2015-09-09
JIANGSU WECARE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In recent years, the application research about the exopolysaccharide of lactic acid bacteria is increasing day by day, utilizes the method that the phosphorylated polysaccharide that produces with Lactococcus lactis etc. is utilized as humectant and whitening agent Publication No.), the method of using polysaccharide components produced by Lactobacillus strains as an anti-inflammatory agent (Japanese Patent Laid-Open Publication No. 7-70209), and the method of using the culture supernatant of lactic acid bacteria containing exopolysaccharide as a wrinkle inhibitor (CN 101505721A); However, there is no report on the use of exopolysaccharides produced by Bifidobacterium longum and Streptococcus thermophilus as skin color improving ingredients

Method used

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  • Extracellular polysaccharide high-yielding lactic acid bacterium BL21, and preparation method and application thereof
  • Extracellular polysaccharide high-yielding lactic acid bacterium BL21, and preparation method and application thereof
  • Extracellular polysaccharide high-yielding lactic acid bacterium BL21, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Isolation, screening and identification of strains

[0029] The feces samples of healthy infants and young children were collected, dissolved in sterile water, serially diluted, added to a sterilized plate, and poured into MRS medium at about 55°C. The formula of MRS medium was: peptone 10.0g, beef extract 8.0 g, yeast extract 4.0g, lactose 20.0g, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, L-cysteine ​​hydrochloride 1.0g, triammonium citrate 2.0 g, Tween 801.0g, agar 20.0g, pH 6.0-6.5, add distilled water to 1000mL, sterilize at 121°C for 20min. Under anaerobic conditions, culture at 37°C for 72h. Select the colonies with large and strong calcium-dissolving circles for streak purification and culture, and then inoculate them into milk with 10% (W / V) solid content at a volume ratio of 1% to 3%, culture at 43°C for 6 hours, and stir the curd and observe the status.

[0030] Select the strains...

Embodiment 2

[0032] Example 2: Fermentation and polysaccharide extraction

[0033] The Bifidobacterium longum BL21 obtained by screening is inoculated into the exopolysaccharide medium, and the percentage by weight of each component in the exopolysaccharide medium: 0.5% fructose, 1.2% sucrose, 0.5% fructooligosaccharide, 0.5% oligosaccharide Galactose was used as a carbon source and the balance of water was first cultured at 37°C for 12 hours, and then the temperature was lowered to 30°C for 36 hours.

[0034] The culture solution was processed with an angle rotor centrifuge, centrifuged at 6000 rpm for 20 minutes, and the first supernatant was collected. Then add the carrageenan aqueous solution of first supernatant weight 2% in the first supernatant liquid, described carrageenan aqueous solution is that the carrageenan after sterilizing is dissolved in water and obtains, and wherein carrageenan content is 0.2wt%, fully After mixing, add 1 / 10 volume of chloroform, oscillate to mix evenly...

Embodiment 3

[0038] Example 3: Effect of exopolysaccharide of Bifidobacterium longum BL21 on melanin formation of B16 cells

[0039] B16 melanoma cells were cultured in DMEM medium containing 5% fetal bovine serum at 37°C and 5% CO 2 Constant temperature culture. Take the B16 melanoma cells in the logarithmic phase, discard the culture medium, wash twice with PBS solution, add the culture medium and pipette, adjust the concentration of the cell suspension to 5×104-10×104 cells / ml, add it to a 96-well plate , add 100 μl to each well. Cultivate for 12-16 hours to allow the cells to adhere to the wall, and aspirate the culture medium.

[0040] The exopolysaccharide obtained in Example 2 was prepared into culture medium with different concentrations, and 10 μl of CCK-8 reagent (CCK-8 Cell Counting Kit, Yeasen) was added at the same time, and the cell viability was tested according to the method specified in the kit. A control group (cells + culture medium) was set, and 5 replicate wells wer...

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Abstract

The invention belongs to the field of microbes, and concretely relates to an extracellular polysaccharide high-yielding lactic acid bacterium, extracellular polysaccharide yielding optimum conditions and a preparation method thereof, an extracellular polysaccharide generated by the bacterium, and a use of the extracellular polysaccharide. The Bifidobacterium longum BL21 provided by the invention is cultured and extracted to obtain the extracellular polysaccharide. The extracellular polysaccharide can be used for preparing external preparations for improving the skin color. The lactic acid bacterium provided by the invention can highly yield the extracellular polysaccharide capable of inhibiting synthesis of melanin in cells and reducing the quantity of skin color spots, the Bifidobacterium longum is a probiotic, and the extracellular polysaccharide produced by fermenting the Bifidobacterium longum is safe, harmless and even beneficial to human bodies and has excellent safety, so the extracellular polysaccharide can be used for preparing cosmetics, skin care products and various skin external use agent compositions.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a lactic acid bacteria BL21 with high production of exopolysaccharides, the exopolysaccharides produced and uses thereof. Background technique [0002] The skin is divided into two parts, the epidermis and the dermis from the outside to the inside, and the depth of the skin color depends on the amount of melanin (also known as melanin) in the epidermis. [0003] Melanin is a group of complex pigment components formed by oxidative polymerization process catalyzed by tyrosinase system with tyrosine as precursor, which can be divided into insoluble black 5,6-dihydroxyindole melanin and soluble relatively High 5,6-dihydroxyindolecarboxylic acid type melanin. Melanin is synthesized in the melanosomes produced by melanocytes in the basal layer of the epidermis and migrates to the surrounding keratinocytes, and continuously migrates to the skin surface with the renewal of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/04A61K8/73A61K31/715A61Q19/02A61Q17/04A61P17/00C12R1/01
Inventor 方曙光马凯姜甜夏九学
Owner JIANGSU WECARE BIOTECHNOLOGY CO LTD
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