Method for screening personalized intestinal environment-improving material and composition therefor
The present disclosure relates to a composition for screening an intestinal environment-improving material and a screening method using the composition, and according to the composition and the method of the present disclosure, it is possible to provide an effective analysis method for screening a microbiota-improving candidate material in a personalized manner by providing a method for verifying personalized probiotics, prebiotics, foods, health functional foods and drugs under in vitro conditions based on microbiota and microbiota metabolites.
1. A method for screening an intestinal environment-improving composition, comprising:
(a) mixing a sample obtained from a subject with medium for screening the intestinal environment-improving composition, and homogenizing the sample, thereby obtaining the sample in medium;
(b) treating the sample in medium from step (a) with the intestinal environment-improving composition and culturing a mixture including the sample in medium and the intestinal environment-improving composition; and
(c) analyzing the cultured mixture from step (b) to determine content or concentration of toxin, intestinal microbiota, microbiota-derived metabolite, or a mixture thereof in the culture thereof,
wherein
the medium for screening the intestinal environment-improving composition does not include protein and carbohydrate; and
the medium for screening the intestinal environment-improving composition comprises L-cysteine.
2. The method of claim 1 , wherein the medium for screening the intestinal environment-improving composition further comprises mucin.
3. The method of claim 1 , wherein the intestinal environment-improving composition is selected from the group consisting of probiotic, prebiotic, food, health functional food, drug and a mixture thereof.
4. The method of claim 1 , wherein the toxin is endotoxin or hydrogen sulfide.
5. The method of claim 1 , wherein the microbiota-derived metabolite is a short-chain fatty acid (SCFA).
6. The method of claim 5 , wherein the short-chain fatty acid is selected from the group consisting of acetate, propionate, butyrate, isobutyrate, valerate and iso-valerate.
7. The method of claim 1 , wherein the method is performed under in vitro conditions.
8. The method of claim 1 , wherein the culturing in step (b) is performed under an anaerobic condition for 18 hours to 48 hours.
9. The method of claim 1 , wherein the analyzing the cultured mixture in the step (c) is to analyze the content, concentration, or type of toxin.
10. The method of claim 1 , wherein the analyzing the cultured mixture in the step (c) is to analyze the kind, concentrate, content or diversity change of intestinal microbiota contained in the culture.
11. The method of claim 5 , further comprising:
(d) screening a candidate composition that increases the content of the short-chain fatty acids, increases the kind and content of beneficial bacteria in the microbiota, decreases the contents of endotoxin and hydrogen sulfide or decreases the kind and content of harmful bacteria in the microbiota by comparison between the result of analysis from the step (c) and the result of analysis on a control group.
12. A method for screening an intestinal environment-improving composition, comprising:
(1) preparing a mixture of human or animal feces and a Pharmaceutical Meta-Analysis Screening (PMAS) medium;
(2) dispensing identical quantities of the feces-PMAS medium mixture into culture plates in an anaerobic chamber;
(3) screening each of candidate intestinal environment-improving compositions;
(4) culturing the prepared plates under anaerobic conditions with temperature, humidity and motion similar to those of the intestinal environment to ferment and culture respective test groups; and
(5) centrifuging the cultured respective test groups of step (4) to obtain supernatant; and
(6) testing for the presence of toxic or undesirable substances comprising hydrogen sulfide, bacterial LPS, short-chain fatty acids in the supernatant, and/or microbiota.
13. The method of claim 12 , wherein step (1) comprises the following steps:
(a) obtaining the human or animal feces and mixing the feces with the PMAS medium,
(b) homogenizing the mixture using a stomacher;
(c) filtering fecal residues from the feces; and
(d) reducing the feces-medium mixture in the anaerobic chamber.
14. The method of claim 12 , wherein step (3) comprises the following steps:
(i) suspending each of the candidate compositions in a sterile buffer; and
(ii) homogenizing each of the candidate compositions and then dispensing the homogenized candidate compositions to the respective plates prepared in Step (2) where the feces-medium mixture was placed.
15. The method of claim 12 , further comprising:
(7) identifying the intestinal environment-improving composition material,
wherein the corresponding cultured sample results in reduced levels of toxic or undesirable substances relative to a control fecal sample not treated with a candidate intestinal environment-improving composition material to screen the intestinal environment-improving composition material.
16. The method of claim 12 wherein the feces and the PMAS medium are mixed in a ratio of 1:12.
17. The method of claim 12 wherein the PMAS medium comprises glucose, tryptone, mucin or L-cysteine hydrochloride.