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A Lactobacillus casei with regulation of antibiotic-induced flora structure disorder

A Lactobacillus casei and microorganism technology, applied in the field of Lactobacillus casei, can solve problems such as blanks, and achieve the effects of long shelf life, alleviation of oxidative stress damage, and high content of active ingredients

Active Publication Date: 2020-02-07
无锡食生臻选生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the disturbance of intestinal flora caused by antibiotics, and the current research on probiotics on the disturbance of intestinal flora caused by antibiotics is relatively blank

Method used

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  • A Lactobacillus casei with regulation of antibiotic-induced flora structure disorder
  • A Lactobacillus casei with regulation of antibiotic-induced flora structure disorder
  • A Lactobacillus casei with regulation of antibiotic-induced flora structure disorder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: The survival rate determination of different Lactobacillus casei in artificial simulated gastrointestinal fluid

[0035] Prepare simulated gastrointestinal juice, simulated gastric juice (6.2g / L NaCl, 2.2g / L KCl, 0.22g / L CaCl 2 ,1.2g / LNaHCO 3 , 0.3% pepsin) was adjusted to pH 2.0 with 6M HCl, and the small intestinal fluid (6.4g / LNaHCO 3, 0.239g / L KCl, 1.28g / L NaCl and 0.1% pancreatin), adjusted to pH 7.4 with 6M HCl. After fully dissolving, the simulated gastric juice and intestinal juice were sterilized by filtration with a 0.22 μm microporous membrane and dispensed into 5 mL in each test tube, and refrigerated at 4°C for later use. Centrifuge the lactic acid bacteria cultured for 18 hours, resuspend and metabolize with 0.9% normal saline for 2 hours, then insert into simulated gastric juice and mix well; draw another 100 μL of gradient dilution plate counting method for counting.

[0036] Set the temperature of the oscillating water bath to 37°C, and...

Embodiment 2

[0042] Embodiment 2: Lactobacillus casei grows in the culture medium that oligosaccharide is the sole carbon source

[0043] Production medium: 4g fructooligosaccharide, 2g tryptone, 1g yeast extract, 2g KH 2 PO 4 , 0.002g MgSO 4 ·7H 2 O, 0.08g NaCl, 8mg CaCl 2 , 0.73mg FeSO 4 ·7H 2 O, 1.2mg hematin, 10mL ATCC vitamin mixture, 10mL ATCC trace mineral, 0.5mL Tween 80, 0.5g L-cysteine ​​hydrochloride, add water to 1000mL, adjust the pH to 7.0, and sterilize at 115°C for 20min. Among them, the fructooligosaccharide is added to the culture medium after being sterilized through a 0.22 μm filter membrane. When separating bacteria, add bromocresol purple solution with a mass concentration of 0.5% as indicator, and the added amount is 15mL / L.

[0044] Use the single carbon source of fructooligosaccharides-bromocresol purple (discoloration 5.7-6.2) solid MRS medium to measure the utilization of probiotics to fructooligosaccharides: growth indicates that oligosaccharides do not i...

Embodiment 3

[0049] Embodiment 3: Comparison of adhesion performance of different Lactobacillus casei to human intestinal cells

[0050] HT-29 cells were cultured in RPMI 1640 (10% fetal bovine serum, 1% antibiotics) medium, and the medium was changed every two days until the cells reached 80-90%. Adjust cell concentration to 1 x 10 5 cells / mL, inoculated in a six-well culture plate, put a 18×18mm sterile coverslip in the culture plate, and place it at 37°C, 95% air / 5% CO 2 Culture in an incubator until a dense monolayer of cells grows. After the cells were washed twice with PBS buffer, 1 mL of antibiotic-free RPMI medium and 1 mL of cultured and resuspended bacteria suspension in PBS were added to each well (the total number of bacteria was 10 8 ), mixed and incubated in an incubator, each strain had three parallels. After culturing for 2 h, the culture plate was taken out, and the cells were washed with PBS buffer until the non-adhered lactic acid bacteria were removed. After adding ...

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Abstract

The invention discloses a Lactobacillus casei capable of regulating the disorder of the flora structure caused by antibiotics, and belongs to the technical field of food. The present invention is to expand the fermentation of the bacterium, add skim milk and trehalose, and freeze-dry to obtain a dietary supplement, which contains more than 10 live bacteria. 9 CFU / g of Lactobacillus casei CGMCC12435. The Lactobacillus casei CGMCC12435 provided by the present invention has good survivability of simulated gastrointestinal tract in vitro, and can grow well by utilizing the culture medium of the sole carbon source of oligosaccharides. High adhesion rate to human HT‑29 cells in vitro. The dietary supplement made from this bacterium can significantly restore the changes in intestinal flora caused by antibiotics in mice, and relieve the oxidative damage caused by antibiotics, and has broad market prospects.

Description

technical field [0001] The invention relates to a Lactobacillus casei capable of regulating the disorder of the flora structure caused by antibiotics, and belongs to the technical field of food. Background technique [0002] Intestinal microorganisms play an important role in human physiological metabolism and avoiding diseases. A large number of symbiotic bacteria live and multiply in the gastrointestinal tract. Process has a crucial impact. The microbiota protects the host from infection by direct defense against invading microorganisms or by coordinating an appropriate immune response. The composition and metabolic activities of human colonic microbes are regulated by a number of external factors, including diet and antibiotic treatment, and changes in the structure and metabolism of gut microbes may have long-term effects on host health. Antibiotics alter microbial composition, which may lead to a series of subsequent symptoms such as increased risk of disease, seconda...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L33/135A23L2/38A23C9/123C12R1/245
CPCA23C9/1234A23L2/382A23L33/135A23V2002/00C12R2001/245C12N1/205A23V2400/125A23V2200/3204A23V2200/30
Inventor 翟齐啸陈卫史瑛田丰伟赵建新张灏
Owner 无锡食生臻选生物科技有限公司
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