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Intestinal simulation model-based lactic acid bacteria intestinal probiotic effect measuring method

A technique of simulating models and measuring methods, which is applied in the field of detection, can solve problems such as difficulty in sampling intestinal bacteria, achieve high accuracy, exclude host factors, and overcome sampling difficulties

Inactive Publication Date: 2019-03-08
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to overcome the shortcomings and deficiencies of the prior art, the object of the present invention is to provide a method for measuring the probiotic effect of lactic acid bacteria on the intestinal tract based on the intestinal simulation model, which overcomes the problem of difficult sampling of intestinal bacteria through in vitro simulated fermentation, and eliminates the host factors

Method used

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  • Intestinal simulation model-based lactic acid bacteria intestinal probiotic effect measuring method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1: Activation of Lactobacillus sake LZ217

[0053] Take out the preserved strain (Lactobacillus sake LZ217) from the refrigerator at -80°C, place it in a refrigerator at 4°C to thaw for 30 minutes, streak on the flat plate MRS solid medium, put it in a 30°C incubator for 36 hours, and pick a single colony to Activated twice in MRS liquid medium to obtain activated Lactobacillus sake LZ217.

[0054] Plate MRS solid medium, made from the following raw materials:

[0055]

[0056]

[0057] First mix the above raw materials, dilute to 1L with water, then adjust the pH to 7.0, then add 15g of agar and sterilize at 121°C for 15min.

[0058] MRS liquid medium, made from the following raw materials:

[0059]

[0060] First mix the above raw materials, dilute to 1 L with water, adjust the pH to 7.0, and then sterilize at 121°C for 15 minutes.

Embodiment 2

[0061] Example 2: In vitro simulation culture of human intestinal flora

[0062] Formulated intestinal simulation medium, made from the following raw materials:

[0063]

[0064]

[0065] First mix the above components, add water to make the volume to 1000mL, and sterilize at 121°C for 15min to obtain intestinal simulated medium.

[0066] (1) Add the activated Lactobacillus sakei LZ217 obtained in Example 1 to the intestinal simulation medium, and add L. sakei LZ217 cells to the experimental group, so that the number of viable L. sakei LZ217 bacteria reaches 1×10 8 CFU / mL (that is, the number of live bacteria of Lactobacillus sake LZ217 in the intestinal simulation medium reaches 1×10 8 CFU / mL), the mixed solution was blown evenly with a pipette gun to obtain an intestinal simulated medium added with Lactobacillus sake LZ217, and without adding Lactobacillus sake LZ217 as a blank control, to obtain an intestinal simulated medium without added Lactobacillus sake LZ217; ...

Embodiment 3

[0070] Embodiment 3: Detection of short-chain fatty acids in fermentation broth by gas chromatography

[0071] (1) Processing of samples. Take out the supernatant obtained in Example 2, filter it with a 0.22 μm filter membrane, take 500 μL of the filtered supernatant and add 100 μL of crotonic acid, shake and mix to obtain a mixed solution, take 20 μL of the mixed solution and inject it into a sample injection bottle detection. All samples to be tested were continuously injected three times on GC (GC series gas chromatograph).

[0072] (2) Column temperature rise program. Column temperature: keep at 70°C for 1min, raise to 180°C at 15°C / min, hold for 2min, raise to 240°C at 40°C / min and hold for 3min; inlet parameters: temperature 250°C, injection volume: 1μL; spectrum Column flow: 2.93mL / min, FID (Flame Ionization Detector) detector parameters: temperature 250°C; gasification chamber: 250°C. Carrier gas: nitrogen, makeup gas flow: 30mL / min, hydrogen flow: 40mL / min, air fl...

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Abstract

The invention discloses an intestinal simulation model-based lactic acid bacteria intestinal probiotic effect measuring method. The intestinal simulation model-based lactic acid bacteria intestinal probiotic effect measuring method comprises the following steps of activating lactobacillus sakei LZ217; adding the activated lactobacillus sakei LZ217 into an intestinal simulation culture medium, meanwhile, setting a control group with no lactobacillus sakei LZ217 added, then adding in processed feces samples for simulating culture to obtain fermentation liquors, performing centrifugal treatment on the fermentation liquors to obtain supernatants; performing gas chromatography analysis on the supernatants to detect short-chain fatty acids. The intestinal simulation model-based lactic acid bacteria intestinal probiotic effect measuring method can be applied to detecting and analyzing the influence of the lactobacillus sakei LZ217 on the intestinal flora change and the short-chain fatty acidmetabolism of the volunteers, and meanwhile, through in vitro simulating fermentation, can overcome the difficulty in sampling intestinal bacteria and eliminate host factors.

Description

technical field [0001] The invention relates to the technical field of detection methods, in particular to a method for measuring the probiotic effect of lactic acid bacteria on the intestinal tract based on an intestinal simulation model. Background technique [0002] Lactic acid bacteria (LAB) is a general term for bacteria that can produce large amounts of lactic acid from fermentable carbohydrates. LAB mainly includes 18 species including Lactococcus, Streptococcus, Enterococcus, Leuconostoc and Lactobacillus. LAB is characterized as nonpathogenic, Gram-positive, catalase-negative, non-spore-forming, facultatively anaerobic, and obligately anaerobic. Lactic acid bacteria can settle in the gastrointestinal tract and exert their beneficial effects for a long time without continuous medical intervention. Due to people's demand for reducing medical costs, lactic acid bacteria have become the most widely used probiotics and play a huge role in the food industry. role. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N1/20G01N30/02C12R1/225
CPCG01N30/02C12N1/20C12Q1/02G01N2333/335
Inventor 顾青郦萍叶开陈青青周青青
Owner ZHEJIANG GONGSHANG UNIVERSITY
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