Bacillus subtilis for improving yield of traditional Chinese medicine polysaccharide and regulating skin barrier and immunity

A technology of Bacillus subtilis and traditional Chinese medicine, applied in the direction of skin diseases, bacteria, drug combination, etc., to reduce NO content and improve cell survival rate

Active Publication Date: 2021-12-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are no strains and fermentation processes that can use Dendrobium and Astragalus raw materials for effective fermentation, and there is no strain or fermentation process that can improve the yield of fermented polysaccharides while improving the efficacy of fermented polysaccharides in skin barrier repair and skin immune regulation

Method used

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  • Bacillus subtilis for improving yield of traditional Chinese medicine polysaccharide and regulating skin barrier and immunity
  • Bacillus subtilis for improving yield of traditional Chinese medicine polysaccharide and regulating skin barrier and immunity
  • Bacillus subtilis for improving yield of traditional Chinese medicine polysaccharide and regulating skin barrier and immunity

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1: Screening and identification of strains

[0065] Specific steps are as follows:

[0066] 1. Screening

[0067] The sample comes from kimchi in Lijiang, Yunnan. After pretreatment, the sample is stored in 20% glycerol in a -80°C refrigerator. After taking out and thawing, mix well and add 0.5mL sample to 4.5mL normal saline to contain 9g / L normal saline for gradient dilution, select the appropriate gradient dilution and spread it on the LB solid medium, culture at 37°C for 48 hours, pick the typical colony of Bacillus subtilis to streak and purify on the LB solid medium, and pick a single colony Transferred to LB liquid medium for enrichment and preserved in 30% glycerol to obtain a strain named CCFM1162; wherein, the typical colony of Bacillus subtilis presents a rough and opaque surface, stained white or yellowish.

[0068] 2. Identification

[0069] The genome of the bacterial strain CCFM1162 was extracted, the 16S rDNA of the bacterial strain CCFM11...

Embodiment 2

[0070] The extraction of embodiment 2 dendrobium crude polysaccharides

[0071] Specific steps are as follows:

[0072] (1) Preparation of Dendrobium raw material fermentation medium: Mix 40g / L Dendrobium raw material, 10g / L yeast extract powder and 10g / L NaCl evenly, add purified water to make up the volume, adjust the pH value to 7.0, and heat at 115°C for 20min to extinguish bacteria;

[0073] (2) Preparation of dendrobium raw material fermentation broth: cool the dendrobium raw material fermentation medium in step (1) to below 40°C, and in a sterile environment, add 1 × 10 to the dendrobium raw material fermentation medium 6 The amount of cfu / mL was added to Bacillus subtilis CCFM1162, and fermented at a constant temperature and pH value of 7.0 at a temperature of 37°C and a pH value of 7.0 for 15-16 hours. At this time, the number of viable bacteria reached ≥10 9 cfu / mL; continue to ferment for 24 hours, reach the end of fermentation, and obtain the raw material ferment...

Embodiment 3

[0077] The extraction of embodiment 3 astragalus crude polysaccharides

[0078] Specific steps are as follows:

[0079] (1) Prepare astragalus raw material fermentation medium: mix 80g / L astragalus raw material, 10g / L yeast extract powder and 10g / LNaCl evenly, add purified water to constant volume, adjust pH value to 7.0, and heat at 115°C for 20min to kill bacteria;

[0080] (2) Preparation of astragalus raw material fermentation broth: cool the sterilized astragalus raw material fermentation medium in step (1) to below 40°C, and in a sterile environment, add 1×10 6 The amount of cfu / mL was added to Bacillus subtilis CCFM1162, and fermented at a constant temperature and pH value of 7.0 at a temperature of 37°C and a pH value of 7.0 for 15-16 hours. At this time, the number of viable bacteria reached ≥10 9 cfu / mL; continue to ferment for 24 hours, reach the end of fermentation, and obtain astragalus raw material fermentation liquid.

[0081] (3) Extracting astragalus polysa...

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Abstract

The invention discloses bacillus subtilis for improving the yield of traditional Chinese medicine polysaccharide and regulating skin barrier and immunity, and belongs to the field of fermentation engineering. Traditional Chinese medicine raw materials are used as a unique carbon source, proliferation factors and microelements beneficial for promoting proliferation of bacillus subtilis are added, the bacillus subtilis can rapidly proliferate in Dendrobium nobile Lindl or radix astragali raw material fermentation liquor, fermentation is conducted for certain time in the mode, the proliferation of the bacillus subtilis in the fermentation liquor exceeds 10 < 9 > cfu/mL, the content of the traditional Chinese medicine polysaccharide extracted after fermentation is increased by 1.13-1.72 times if being compared with the extraction rate of the traditional Chinese medicine polysaccharide extracted without fermentation, and the content of the traditional Chinese medicine polysaccharide extracted after fermentation is remarkably increased if being compared with the unfermented polysaccharide on the aspects of skin barrier repair and immunoregulation, so that the cell survival rate of SDS-induced HaCaT is remarkably increased, and the NO content in LPS-induced cells is reduced.

Description

technical field [0001] The invention relates to bacillus subtilis which improves the yield of traditional Chinese medicine polysaccharide and regulates skin barrier and immunity, and belongs to the field of fermentation engineering. Background technique [0002] Dendrobium is a famous and precious traditional Chinese medicine in my country, known as the "gold in medicine". Dendrobium is sweet in taste and slightly cold in nature. It has the effects of nourishing the stomach and promoting body fluid, nourishing yin and clearing away heat. Studies have shown that the main active ingredients contained in Dendrobium are Dendrobium polysaccharides, dendrobine, amino acids, stilbenes and their derivatives, and various volatile components. Polysaccharides are the most important active ingredients. The content of polysaccharides determines the biological activity of Dendrobium. Strong and weak, it has the functions of enhancing immunity, anti-fatigue, anti-oxidation, lowering blood...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K31/715A61P17/16A61P37/02C12P19/04C12R1/125
CPCC12P19/04A61K31/715A61P17/16A61P37/02Y02A50/30
Inventor 崔树茂唐鑫毛丙永赵建新
Owner JIANGNAN UNIV
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