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Method for evaluating activity of dietary polysaccharide

A polysaccharide and diet technology, applied in the direction of biochemical equipment and methods, microbial measurement/testing, instruments, etc., can solve the problem that the biological activity of dietary polysaccharides cannot be accurately reflected

Inactive Publication Date: 2020-10-23
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the use of cell models to evaluate the activity of polysaccharides mainly involves the direct intervention of dietary polysaccharide solutions in cells, which cannot accurately reflect the biological activity of dietary polysaccharides.

Method used

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  • Method for evaluating activity of dietary polysaccharide
  • Method for evaluating activity of dietary polysaccharide
  • Method for evaluating activity of dietary polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Replace the carbon source glucose in the microbial basal medium with tea polysaccharide, that is, the medium contains: tea polysaccharide 10g / L, peptone 2.0g / L, yeast extract 2.0g / L, sodium chloride 0.1g / L, phosphoric acid Dipotassium hydrogen 0.04g / L, potassium dihydrogen phosphate 0.04g / L, magnesium sulfate 0.01g / L, calcium chloride 0.01g / L, sodium bicarbonate 2.0g / L, hemin 0.02g / L, semi Cystine hydrochloride 0.5g / L, bile salt 0.5g / L, resazurin 1.0g / L, Tween 80 2.0mL / L and vitamin K1 10μL / L;

[0020] (2) Fermented stool samples in vitro were provided by 4 healthy volunteers (2 males and 2 females, aged 22 to 28 years old). The volunteers had not taken antibiotics or probiotics within 3 months and had no gastrointestinal diseases. Immediately after each volunteer collected 5g of fresh feces, mixed with 45g of normal saline (NaCl 8.5g / L, cysteine ​​hydrochloride 0.5g / L), stirred evenly, and centrifuged (250rmp) to obtain a 10% feces suspension;

[0021] (3) Take 2 ...

Embodiment 2

[0026] (1) Replace the carbon source glucose in the microbial basal medium with mycopolysaccharide, that is, the medium contains: 10g / L of mycopolysaccharide, 2.0g / L of peptone, 2.0g / L of yeast extract, and 0.1g of sodium chloride / L, dipotassium hydrogen phosphate 0.04g / L, potassium dihydrogen phosphate 0.04g / L, magnesium sulfate 0.01g / L, calcium chloride 0.01g / L, sodium bicarbonate 2.0g / L, hemin 0.02g / L, cysteine ​​hydrochloride 0.5g / L, bile salt 0.5g / L, resazurin 1.0g / L, Tween 80 2.0mL / L and vitamin K1 10μL / L;

[0027] (2) Fermented stool samples in vitro were provided by 4 healthy volunteers (2 males and 2 females, aged 22 to 28 years old). The volunteers had not taken antibiotics or probiotics within 3 months and had no gastrointestinal diseases. Immediately after each volunteer collected 5g of fresh feces, mixed with 45g of normal saline (NaCl 8.5g / L, cysteine ​​hydrochloride 0.5g / L), stirred evenly, and centrifuged (250rmp) to obtain a 10% feces suspension;

[0028] (...

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Abstract

The invention relates to a method for evaluating the activity of dietary polysaccharide. The method comprises the following steps: adding dietary polysaccharide into a microbial culture medium as a carbon source, culturing the dietary polysaccharide and fecal microorganisms at 37 DEG C for 24 hours under an in-vitro anaerobic condition, performing centrifuging at 10000rmp, passing through a 0.22 [mu]m membrane, collecting fermentation liquor, and detecting metabolites of the dietary polysaccharide by using metabonomics. Caco-2 cells are planted in an upper chamber and Caco-2 cells for evaluating the activity of the dietary polysaccharide are planted in a lower chamber by using a Transwell technology; after the fermentation liquor is added into an upper chamber cell culture medium to be cultured for 24 h, liquid phase, gas phase or metabonomics and other means are used for detecting which metabolites can penetrate through a Caco-2 monolayer cell model, and related indexes of lower chamber cells are measured. The method provided by the invention can be used for systematically and scientifically evaluating the biological activity of the dietary polysaccharide.

Description

technical field [0001] The invention relates to the field of food science and engineering, in particular to a method for evaluating the activity of dietary polysaccharides. Background technique [0002] Polysaccharides are natural polymer compounds in higher plants, animal cell membranes, and microbial cell walls, and are widely involved in various life phenomena of cells. Due to its diverse biological activities and wide use in the fields of food health care and disease prevention, the research on the development and utilization of polysaccharide resources has become increasingly active, and has become a research hotspot in food science, natural medicine and biochemistry. More and more studies have shown that dietary polysaccharides have functions such as anti-oxidation, antibacterial, anti-obesity, anti-inflammation, anti-diabetes, anti-tumor, protection of cardiovascular and nervous systems, and reduction of UV damage to the skin. At present, there are more than 300 kind...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N33/68
CPCG01N33/5008G01N33/5038G01N33/6866G01N33/6869G01N2333/56G01N2333/5412
Inventor 陈贵杰曾晓雄孙怡
Owner NANJING AGRICULTURAL UNIVERSITY
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