JP2006314219A - Method for adsorbing bile acid utilizing enterobacterium, and bile acid-adsorbing substance - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for suppressing biosyntheses or promoting metabolism owing to reducing in vivo cholesterol levels as a result of increasing bile acid secretion by bile acid intake. <P>SOLUTION: Owing to enterobacteria having such characteristics as to positively make an intake of bile acid in the presence of a saccharide source such as glucose and release no bile acid extracellularly while the saccharide source is supplied, and since these enterobacteria exist predominantly in the intestine and form no secondary bile acid, the formation of such harmful secondary bile acid concomitant with the increase in bile acid secretion can also be suppressed. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

The present invention relates to a bile acid adsorption method and a bile acid adsorption substance using an intestinal bacterium having a property of taking bile acid into cells.

In recent years, the number of sufferers of hypercholesterolemia is increasing with the westernization of dietary habits. Hypercholesterolemia is a major risk factor for arteriosclerosis, but its prevention and improvement requires major changes in lifestyle such as diet and exercise, and sometimes it is forced to maintain self-regulation that is difficult to sustain . A simple preventive / improvement measure that anyone can accept is desired.

Examples of methods for reducing cholesterol in vivo include a method for suppressing absorption of food-derived cholesterol, a method for suppressing in vivo cholesterol biosynthesis, and a method for promoting metabolism of in vivo cholesterol. Since most of the in vivo cholesterol is biosynthesized in the body, the method of suppressing biosynthesis or promoting metabolism is considered to contribute more to cholesterol reduction than the method of suppressing absorption.

In vivo cholesterol is metabolized to bile acids, steroid hormones, etc., and the most quantitatively is metabolism to bile acids. Bile acids are synthesized in the liver using cholesterol as a raw material and then secreted as a glycine or taurine conjugate in the upper small intestine to promote digestion and absorption of lipids in foods. Most of the bile acids are actively reabsorbed in the lower part of the small intestine, but some of them enter the large intestine and become free bile acids by intestinal bacteria having a deconjugating action. It is converted into secondary bile acids such as deoxycholic acid by bacteria. Secondary bile acids are known to act as promoters for colorectal cancer.

In the human intestinal tract, there are about 10 ¹¹ -10 ¹² , about 500 types of intestinal bacteria per gram of feces. Among the enterobacteria, useful functions such as lactic acid bacteria have been reported for many functions such as cholesterol reduction, but the ratio of these to the whole intestinal flora is about several percent at the maximum. The remaining bacterial groups such as the Collinsella genus and Clostridium genus are called opportunistic bacteria, and despite the overwhelming number of bacteria, there are many unclear points regarding their functions.

By suppressing the reabsorption of bile acids as described above, it is possible to reduce cholesterol in the living body as a raw material. The detailed method is as follows. 1. Promotion of deconjugation 2. transformation or decomposition; Simple adsorption, 4. For example, uptake into cells.
1. Is a method that utilizes the fact that free bile acids are less susceptible to active reabsorption in the small intestine, such as a method that utilizes a conjugated bile acid deconjugating enzyme protein (see Patent Document 1). However, since free bile acids are substrates for 7α dehydration oxidation, if deconjugation is promoted by this method, the amount of harmful secondary bile acids may increase and adversely affect the living body. It's not a good way.

2. Is a method of reducing the amount of reabsorption by converting or decomposing bile acids in the intestinal tract, and examples thereof include lactic acid bacteria listed in Patent Document 2. However, conversion of bile acids may produce new harmful secondary bile acids, and this method is also not sufficient.
3. Is a method of increasing the amount of excretion by simply adsorbing bile acids to resin, dietary fiber, lactic acid bacteria, etc., and the mechanism of action of cholestyramine, a therapeutic drug for hyperlipidemia. Examples of food include proteolytic substances listed in Patent Document 3 and functional foods and drinks containing Lactobacillus casei listed in Patent Document 4. However, since bile acids are exposed to the outside in a simple adsorption state, 7α-dehydrated oxidative bacteria can come into contact with bile acids, so that harmful secondary bile acids may be generated. . 2. Is not enough as well.

4). Is a method of increasing the amount of excretion by incorporating bile acids into the microbial cells in the presence of a sugar source, and there are lactic acid bacteria listed in Patent Document 5. According to this method, bile acids are excreted in the state of being taken up into the microbial cells. To 3. The increase in the amount of secondary bile acid that was concerned by this method does not occur. However, as described above, lactic acid bacteria are an extremely small group of bacteria when viewed from the whole intestinal bacteria, and Patent Documents 5 and 6 are insufficient in terms of the number of bacteria having bile acid uptake ability.

JP 2000-166548 A JP 2004-208577 A JP 2004-203859 A JP 2000-197469 A JP 2001-97870 A International Publication No. 03/013559 Pamphlet

An object of the present invention is to provide a method for adsorbing bile acids by using a bacterium having a property of taking bile acids into the cells, and a bile acid adsorbing substance. More specifically, the present invention relates to a technique for increasing the amount of bile acid excretion without increasing the amount of secondary bile acid and reducing in vivo cholesterol using intestinal bacteria present in large quantities in the intestine. .

As a result of intensive studies on the above-mentioned problems, the present inventors have taken in bile acids actively in the presence of a sugar source such as glucose, and do not release bile acids outside the cells while the sugar source is being supplied. The intestinal bacterium which has the property and exists predominantly in the intestine was found. Furthermore, they found that these intestinal bacteria do not produce secondary bile acids, and completed the present invention.
That is, the present invention provides a method for adsorbing bile acids, characterized by using an enteric bacterium having a property of taking bile acids into the cells in the presence of a sugar source. The present invention also relates to a bile acid adsorbing substance characterized by comprising an intestinal bacterium having a property of taking bile acids into the cells in the presence of a sugar source, and taking bile acids into the cells in the presence of a sugar source. A bile acid adsorbing substance characterized by comprising an assimilating saccharide of enteric bacteria having properties, an enteric bacterium having a property of taking bile acid into the microbial cells in the presence of a sugar source, and a combination of such assimilating sugars A bile acid-adsorbing material is provided.

As the above-mentioned adsorption mechanism, intestinal bacteria having the property of taking bile acids into the cells in the presence of a sugar source are much more present than lactic acid bacteria having similar properties. Compared with the bile acid adsorbent or bile acid adsorption method used, it is considered that this results in stronger bile acid excretion and reduces in vivo cholesterol. Moreover, since these intestinal bacteria do not produce secondary bile acids, production of harmful secondary bile acids associated with an increase in bile acid excretion, which was a problem of the prior art, can also be suppressed.

Examples of enterobacteria that can be used in the present invention include Collinsella, Clostridium, Ruminococcus, and Bacteroides. Specifically, among the strains registered and stored in the Microbiological Materials Development Department (JCM) of the RIKEN BioResource Center, the Collinsella genus includes Collinsella aerofaciens JCM7791, Collinsella aerofaciens JCM10188 strain, Collinsella aerofaciens JCM10789 strain, Collinsella stercoris JCM10709 strain, etc. can be used, and Clostridium spp. Use Clostridium clostridiiforme JCM1291, Strastridium innocuum JCM1292, Clostridium ramosum JCM1298, Clostridium butyricum JCM1391, etc. As the genus Ruminococcus, the Ruminococcus productus JCM1471 strain can be used, and the Bacteroides genus Bacteroides distasonis JCM5825, Bacteroides vulgatus JCM5826 strain, Bacteroides stercoris JCM9496 strain and the like can be used.

As the assimilating sugar used in the present invention, any sugar, oligosaccharide, or polysaccharide can be used as long as it is a sugar that is usually used in foods. Examples of monosaccharides include L-arabinose, ribose, D-xylose, galactose, glucose, fructose, mannose, sorbose, rhamnose, D-lyxose, etc., and disaccharides include cellobiose, maltose, lactose, melibiose, Examples include sucrose, trehalose, gentibiose, D-turanose, lactulose, etc., trisaccharides include melezitose, raffinose, etc., and polysaccharides include inulin, starch, glycogen, fructooligosaccharide, galactooligosaccharide, xylo-oligosaccharide, iso Examples include maltooligosaccharides, gentio-oligosaccharides, and yeast mannan. Examples of sugar alcohols include adonitol, D-arabitol, glycerol, dulcitol, mannitol, sorbitol, and the like. Other sugars analogs, alpha-methyl -D- glucoside, N- acetyl glucosamine, amygdalin, arbutin, esculin, salicin, and the like. Preferably, L-arabinose, ribose, D-xylose, galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, gentibiose, D-turanose, lactulose, raffinose, starch, glycogen, fructooligosaccharide, Examples include galactooligosaccharide, xylo-oligosaccharide, isomaltoligosaccharide, gentio-oligosaccharide, yeast mannan, α-methyl-D-glucoside, N-acetylglucosamine, amygdalin, arbutin, esculin, and salicin.

In addition, an indigestible or low absorbable sugar can be used as appropriate. For example, L-arabinose, D-xylose and the like are listed as monosaccharides, cellobiose, melibiose, gentibiose, lactulose and the like are listed as disaccharides, and raffinose and the like are listed as trisaccharides. , Inulin, fructooligosaccharide, galactooligosaccharide, xylo-oligosaccharide, isomaltoligosaccharide, gentio-oligosaccharide, yeast mannan and the like, and sugar alcohols include dulcitol and sorbitol.
In addition, these saccharides can be used alone, but a plurality of saccharides can be used in combination depending on the application.

Among the above-mentioned sugars, a method for selecting a sugar corresponding to an assimilation sugar of the bile acid uptake enterobacteria is to add a test substance to the bile acid uptake enterobacteria and to bile acid uptake bile acids of the enterobacteria Examples include a method of selecting a test substance that induces an uptake reaction. For example, a bacterial suspension is made to have a McFarland turbidity of 1 with respect to a BCP-added PY medium containing 0.5% of each saccharide alone. There is a method of inoculating, anaerobic culture at 37 ° C. for 12 hours, and evaluating by discoloration of the indicator bromocresol purple.
For example, assimilation sugars for Clostridium bacteria include L-arabinose, ribose, D-xylose, galactose, glucose, fructose, mannose, rhamnose, dalitol, sorbitol, α-methyl-D-glucoside, N-acetylglucosamine, amygdalin , Arbutin, esculin, salicin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, raffinose, starch, glycogen, gentiobiose, D-turanose, D-lyxose, D-arabitol, fructooligosaccharide, galactooligosaccharide, xylooligosaccharide, isomaltoligo Examples include sugars, lactulose, gentio-oligosaccharides, and yeast mannan. Sucrose, fructose, mannose, salicin, cellobiose, maltose, lactose, sucrose, trehalose, etc., and assimilating sugars against luminococcus are glycerol, L-arabinose, ribose, D-xylose, adonitol, galactose, glucose , Fructose, mannose, sorbose, rhamnose, mannitol, sorbitol, amygdalin, esculin, salicin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, inulin, raffinose, starch, etc. , L-arabinose, ribose, D-xylose, galactose, glucose, fructose, rhamnose, amygdalin, esculin, sari Syn, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, inulin, raffinose, starch and the like.

Further, since bile acid uptake enterobacteria exert their effects mainly in the large intestine, the assimilated sugar used in the present invention is an indigestible or low absorbable sugar that reaches the large intestine without being decomposed or absorbed in the small intestine. preferable. For example, low-absorbable sugars include L-arabinose, D-xylose, dulcitol, mannitol, sorbitol, etc., and as non-digestible sugars, cellobiose, melibiose, inulin, raffinose, gentibiose, fructooligosaccharide, galactooligosaccharide, Examples include xylooligosaccharide, isomaltoligosaccharide, lactulose, gentio-oligosaccharide, and yeast mannan. Furthermore, even if the sugar is decomposed and absorbed in the small intestine by utilizing a drug delivery system (DDS) or the like that can reach the large intestine without dissolving the bile acid uptake intestinal bacterial assimilation sugar in the small intestine. It can be used sufficiently.

These enterobacteria can be obtained, for example, by screening shown in the following (1) to (3).
(1) Indirect bile acid uptake test After intestinal bacteria are cultured, they are collected by centrifugation, and the collected bacteria are suspended in a potassium phosphate buffer or the like. A sugar source such as bile acid and glucose is added to the suspension and incubated, and the supernatant is collected by centrifugation at regular intervals, and the bile acid concentration is measured. A strain having a reduced bile acid concentration in the supernatant is selected as compared with the case where no sugar source was added.
(2) Direct bile acid uptake test After intestinal bacteria are cultured, they are collected by centrifugation, and the collected bacteria are suspended in a potassium phosphate buffer or the like. Add a sugar source such as bile acid and glucose labeled with a radioactive isotope such as ¹⁴ C to the suspension, incubate, collect the cells by filtration at regular intervals, and measure the radioactivity of the cells . Compared to the case where no sugar source was added, a strain having a higher radioactivity of the bacterial cells is selected.

(3) Secondary bile acid production test After intestinal bacteria are cultured, they are collected by centrifugation, and the collected bacteria are suspended in a potassium phosphate buffer or the like. Bile acid is added to the suspension and incubated, and the bile acid fraction in the supernatant is extracted, and the bile acid composition is analyzed by thin layer chromatography or the like. A strain in which spots other than bile acids as a substrate were not detected is selected.
The above tests (1) and (2) both measure bile acid uptake activity, and either can be omitted.
As the bile acid used in the tests (1) to (3), various known bile acids can be used, and cholic acid can be particularly preferably used.
In addition, since it is thought that it is mainly in the large intestine that enterobacteria exert their effects, in the test of (1), (2) above, a strain showing a high uptake activity near neutral pH close to the colon pH, It is desirable to select a strain that does not show secondary bile acid production in the tests of (3).

According to the present invention, the amount of bile acid excreted can be increased and the in vivo cholesterol can be reduced by incorporating bile acid into intestinal bacteria in the intestine.
More specifically, the amount of bile acid excretion can be increased by using an intestinal bacterium that actively takes in bile acids in the presence of a sugar source and does not produce secondary bile acids. Furthermore, since no harmful secondary bile acids are produced, it is possible to safely reduce in vivo cholesterol.

In the present invention, a bile acid is actively taken in the presence of a sugar source, and the bile acid is not released outside the cells while the sugar source is supplied, and a secondary bile acid is generated. Use enterobacteria that do not.
As an embodiment of the present invention, for example, there are preparations and foods and drinks in which enterobacteria having the above properties are contained as bacterial cells or cultures containing bacterial cells. By ingesting these preparations and foods and drinks, enteric bacteria having an uptake ability can reach the intestine and the amount of bile acids in the intestine can be reduced.
Further, for example, there are preparations and foods and drinks containing an assimilating sugar that can be used as a sugar source for enteric bacteria having the above properties. By ingesting these preparations and foods and drinks, it is possible to induce the bile acid uptake activity of intestinal bacteria having the uptake ability originally present in the intestine, and to reduce the amount of bile acids in the intestine.

In addition, there are, for example, enterobacteria having the above-mentioned properties or cultures containing the cells, and preparations and foods and drinks containing an enterobacterial saccharide having the above-mentioned properties. By ingesting these preparations and foods and drinks, intestinal bacteria that have the ability to take up bile acids can reach the intestine and induce their bile acid uptake activity to more effectively reduce the amount of bile acids in the intestines. can do.
Examples of the above preparation include powders, granules, capsules, tablets and the like. Examples of the food and drink include various items such as fermented milk, dairy products, beverages, confectionery, and breads.
The amount of enterobacteria having the above properties used in the present invention or the culture containing the cells is 1 × 10 ⁴ to 1 × 10 ¹¹ as a total dose per day, preferably 1 × 10 ⁵ to 1 × 10 ¹⁰ pieces.
Since the assimilating saccharide of the intestinal bacterium having the above properties used in the present invention has no problem in safety, there is no limitation on the dose when it is orally administered. Generally, it is set to the dosage used for food, and specifically, the total dosage per day is 0.05 to 25 g, preferably 0.1 to 20 g. However, for indigestible sugar, the daily dose is preferably less than 10 g.
The blending ratio of the enterobacterial cells having the above-mentioned properties used in the present invention or a culture containing the cells and the assimilating sugars of the intestinal bacteria having the above-mentioned properties is within the range of the daily dose. preferable.
Hereinafter, the present invention will be described more specifically with reference to examples.

Tests for assimilation of bile acid uptake enterobacteria Bacterial assimilation tests were conducted on enterobacteria. The BCP-added PY medium containing 0.5% of each saccharide shown in Table 1 alone was inoculated with the bacterial suspension so that the McFarland turbidity was 1. Anaerobic culture was performed at 37 ° C. for 12 hours, and a sugar in which discoloration of bromocresol purple as an indicator was observed was used as an assimilating sugar. The medium composition is shown below.
BCP-added PY medium (1 liter)
Polypeptone 10g
Yeast extract 5g
Tween 80 1mL
Dipotassium hydrogen phosphate 2g
Sodium acetate trihydrate 5g
2g diammonium citrate
Magnesium sulfate heptahydrate 0.20g
Manganese sulfate tetrahydrate 0.05g
Bromocresol purple 0.17g
Table 1 shows the results of a glucose utilization test using Clostridium butyricum JCM1391 strain. The JCM1391 strain is L-arabinose, ribose, D-xylose, galactose, glucose, fructose, mannose, α-methyl-D-glucoside, N-acetylglucosamine, amygdalin, arbutin, esculin, salicin, cellobiose, maltose, lactose, melibiose, Sucrose, trehalose, raffinose, starch, glycogen, gentiobiose, D-turanose, fructooligosaccharide, galactooligosaccharide, xylooligosaccharide, isomaltoligosaccharide, lactulose, gentiooligosaccharide, and yeast mannan were assimilated.

Measurement of bile acid uptake activity of enterobacteria Indirectly the dominant human enterobacteria, Collinsella, Clostridium, Ruminococcus, Bacteroides Bile acid uptake test was performed.
Enterobacteriaceae were cultured overnight in a commercially available GAM medium. The medium composition is as follows.
GAM medium (1 liter)
Peptone 10g
Proteose peptone 10g
Soy peptone 3g
5g soluble starch
Glucose 3g
Cysteine hydrochloride 0.3g
Yeast extract 5g
Liver extract powder 1.2g
Meat extract powder 2.2g
Digested blood 13.5g
Sodium thioglycolate 0.3g
Sodium dihydrogen phosphate 2.5g

The cells cultured overnight were replanted in the same medium, and the cells in the logarithmic growth phase were collected by centrifugation. The cells were washed once with a potassium phosphate buffer (150 mM potassium phosphate, 1 mM magnesium sulfate, pH 7.0), suspended in the same buffer, and adjusted so that the turbidity at 660 nm was 20. 0.25 mL of 2 mM sodium cholate was added to 4.75 mL of this bacterial cell suspension and allowed to stand at 37 ° C. for 20 minutes, and then 50 μL of 1 M glucose was added to initiate the bile acid uptake reaction. Collect 0.5 mL of the reaction solution over time between 0 and 30 minutes after the start of the reaction, quickly separate the supernatant by centrifugation, and measure the bile acid concentration in the supernatant using the total bile acid test Wako (Wako Pure Chemical Industries, Ltd.) Measured by Kogyo Co., Ltd.). As a control, the concentration of bile acid in the supernatant was measured in the same manner for the sample to which glucose was not added. The decrease in bile acid concentration due to the addition of glucose was taken as the amount of bile acid taken up by the bacteria.

FIG. 1 shows the bile acid uptake result of Clostridium butyricum JCM1391 strain. By adding glucose, cholic acid was quickly taken up into the cells and gradually released after 10 minutes of the reaction that consumed glucose. Similarly, all the test bacteria were tested, and the results are summarized in Table 3. Bile acid uptake activity was observed in the genus Collinsella, Clostridium, Ruminococcus, and Bacteroides.

In the table, T indicates a standard strain.
A direct bile acid uptake test was performed on the strains showing strong bile acid uptake activity for confirmation. A cell suspension was prepared as described above, 50 mM of 2 mM sodium cholate (cholic acid labeled with ¹⁴ C, 197 MBq / mmol) was added to 950 μL of this cell suspension, and the mixture was allowed to stand at 37 ° C. for 20 minutes. Thereafter, 10 μL of 1M glucose was added to initiate the bile acid uptake reaction. From 0 to 30 minutes after the start of the reaction, 100 μL of the reaction solution was collected over time, filtered through a cellulose acetate membrane to separate the cells, and the radioactivity was measured with a liquid scintillation counter. The amount was calculated. As a control, the radioactivity was also measured in the same manner with no glucose added. As an example, the results of Clostridium butyricum JCM1391 strain are shown in FIG. 2, and the same results as in the indirect bile acid uptake test were obtained.

Intestinal Bacterial Bile Acid Uptake by Assimilating Sugar The strains that showed bile acid uptake activity in Example 1 were subjected to an indirect bile acid uptake test in which sugars other than glucose were added. As an example, the test results of Clostridium butyricum JCM1391 strain are summarized in Table 4. Assimilation sugars raffinose, fructooligosaccharides, xylo-oligosaccharides, isomaltoligosaccharides, gentio-oligosaccharides, galactooligosaccharides all induce bile acid uptake like glucose, and raffinose, gentio-oligosaccharides, galacto-oligosaccharides in particular uptake stronger than glucose Inducible activity was shown.

Investigation of secondary bile acid production activity of enterobacteria The secondary bile acid production activity of the strain that showed bile acid uptake activity in Example 2 was investigated. Prepare a cell suspension as described above, add 50 μL of 2 mM sodium cholate to 95 μL of the cell suspension, let stand at 37 ° C. for 20 minutes, and then add 5 μL of 1 M glucose to react with bile acid uptake. Started. 30 minutes after the start of the reaction, 10 μL of 6N hydrochloric acid was added to stop the reaction, and the reaction solution was acidified. The reaction solution was extracted twice with 5 times the amount of ethyl acetate, concentrated and then spotted on a silica gel plate (Silica gel 60, Merck). The silica gel plate is developed with a developing solvent (ethyl acetate: cyclohexane: acetic acid = 69: 21: 9), sprayed with 10% molybdophosphoric acid-ethanol solution and heated at 110 ° C. for 10 minutes to confirm a bile acid blue spot did.
Bile acid spots other than cholic acid added first were not detected, and it was found that the strain showing bile acid uptake activity in Example 1 did not have secondary bile acid production activity. As an example, the results of Clostridium butyricum JCM1391 strain are shown in FIG.

Preparation of Enteric Bacteria and Cultures Containing Bacteria Intestinal bacteria were anaerobically cultured at 37 ° C. for 24 hours in a commercially available GAM medium to obtain a culture solution. The culture was collected as it was or by centrifugation and freeze-dried to obtain a culture and cells containing enterobacteria cells. In Clostridium butyricum JCM1391 strain, 2.0 × 10 ⁷ cells were obtained per 1 mL of the medium.

Tablets, capsules Fungus body of Example 5 10.0 g
Lactose 75.0g
Magnesium stearate 15.0g
Total 100.0g
The above respective parts by weight were uniformly mixed, and tablets and capsules were prepared according to a conventional method.

Tablets, capsules Raffinose 10.0g
Lactose 75.0g
Magnesium stearate 15.0g
Total 100.0g
The above respective parts by weight were uniformly mixed, and tablets and capsules were prepared according to a conventional method.

Tablets, capsules, bacterial cells of Example 5 5.0 g
Raffinose 5.0g
Lactose 75.0g
Magnesium stearate 15.0g
Total 100.0g
The above respective parts by weight were uniformly mixed, and tablets and capsules were prepared according to a conventional method.

Tablets, capsules 10.0 g of culture containing the cells of Example 5
Lactose 75.0g
Magnesium stearate 15.0g
Total 100.0g
The above respective parts by weight were uniformly mixed, and tablets and capsules were prepared according to a conventional method.

Tablets and capsules 5.0 g of the culture containing the cells of Example 5
Raffinose 5.0g
Lactose 75.0g
Magnesium stearate 15.0g
Total 100.0g
The above respective parts by weight were uniformly mixed, and tablets and capsules were prepared according to a conventional method.

Powder, granule 20.0 g of the cells of Example 5
Starch 30.0g
Lactose 50.0g
Total 100.0g
Each said weight part was mixed uniformly, and it was set as the powder and the granule according to the conventional method.

Powder, granule Raffinose 20.0g
Starch 30.0g
Lactose 50.0g
Total 100.0g
Each said weight part was mixed uniformly, and it was set as the powder and the granule according to the conventional method.

Powder, granule Example 5 cells 10.0g
Raffinose 10.0g
Starch 30.0g
Lactose 50.0g
Total 100.0g
Each said weight part was mixed uniformly, and it was set as the powder and the granule according to the conventional method.

Powder, granule 20.0 g of the culture containing the cells of Example 5
Starch 30.0g
Lactose 50.0g
Total 100.0g
Each said weight part was mixed uniformly, and it was set as the powder and the granule according to the conventional method.

Powder, granule 10.0 g of culture containing the cells of Example 5
Raffinose 10.0g
Starch 30.0g
Lactose 50.0g
Total 100.0g
Each said weight part was mixed uniformly, and it was set as the powder and the granule according to the conventional method.

０．０ Sucrose 20.0g
Chickenpox (75% solids) 70.0g
9.5g of water
Perfume 0.45g
Coloring agent 0.045g
0.005 g of the bacterial cells of Example 5
Total 100.0g
Using each component of each of the above parts by weight, it was made into a candy according to a conventional method.

飴 Raffinose 5.0g
Sucrose 15.0 g
Chickenpox (75% solids) 70.0g
9.5g of water
Perfume 0.45g
Coloring agent 0.05g
Total 100.0g
Using each component of each of the above parts by weight, it was made into a bowl according to a conventional method.

飴 Raffinose 5.0g
Sucrose 15.0 g
Chickenpox (75% solids) 70.0g
9.5g of water
Perfume 0.45g
Coloring agent 0.045g
0.005 g of the bacterial cells of Example 5
Total 100.0g
Using each component of each of the above parts by weight, it was made into a candy according to a conventional method.

０．０ Sucrose 20.0g
Chickenpox (75% solids) 70.0g
9.5g of water
Perfume 0.45g
Coloring agent 0.045g
0.005 g of the culture containing the cells of Example 5
Total 100.0g
Using each component of each of the above parts by weight, it was made into a candy according to a conventional method.

飴 Raffinose 5.0g
Sucrose 15.0 g
Chickenpox (75% solids) 70.0g
9.5g of water
Perfume 0.45g
Coloring agent 0.045g
0.005 g of the culture containing the cells of Example 5
Total 100.0g
Using each component of each of the above parts by weight, it was made into a candy according to a conventional method.

Juice Concentrated tangerine juice 15.0g
Fructose 5.0g
Citric acid 0.2g
Fragrance 0.1g
0.15 g of color
Sodium ascorbate 0.048g
0.002 g of the bacterial cells of Example 5
79.5g of water
Total 100.0g
Using each component of the above-mentioned parts by weight, juice was prepared according to a conventional method.

Juice raffinose 2.5g
Concentrated tangerine juice 15.0g
Fructose 2.5g
Citric acid 0.2g
Fragrance 0.1g
0.15 g of color
Sodium ascorbate 0.05g
79.5g of water
Total 100.0g
Using each component of the above-mentioned parts by weight, juice was prepared according to a conventional method.

Juice raffinose 2.5g
Concentrated tangerine juice 15.0g
Fructose 2.5g
Citric acid 0.2g
Fragrance 0.1g
0.15 g of color
Sodium ascorbate 0.048g
0.002 g of the bacterial cells of Example 5
79.5g of water
Total 100.0g
Using each component of the above-mentioned parts by weight, juice was prepared according to a conventional method.

Juice Concentrated tangerine juice 15.0g
Fructose 5.0g
Citric acid 0.2g
Fragrance 0.1g
0.15 g of color
Sodium ascorbate 0.048g
0.002 g of culture containing the cells of Example 5
79.5g of water
Total 100.0g

Juice raffinose 2.5g
Concentrated tangerine juice 15.0g
Fructose 2.5g
Citric acid 0.2g
Fragrance 0.1g
0.15 g of color
Sodium ascorbate 0.048g
0.002 g of culture containing the cells of Example 5
79.5g of water
Total 100.0g
Using each component of the above-mentioned parts by weight, juice was prepared according to a conventional method.

FIG. 4 is a diagram showing the results of an indirect bile acid uptake test of Clostridium butyricum JCM1391 strain. It is a figure which shows the result of the direct bile acid uptake | capture test of JCM1391 strain | stump | stock. It is a development view of a silica gel plate showing a bile acid spot by a standard substance and a JCM1391 strain reaction supernatant extract.

Claims (21)

A method for adsorbing bile acids, comprising using an intestinal bacterium having a property of taking bile acids into cells in the presence of a sugar source. The method according to claim 1, wherein the intestinal bacterium is a Clostridium genus, a Collinsella genus, a Ruminococcus genus, or a Bacteroides genus. The method according to claim 1, wherein the intestinal bacterium is a bacterium having no secondary bile acid producing activity. The method according to claim 1, wherein the intestinal bacterium is Clostridium butyricum. The method according to claim 1, wherein the sugar source is glucose, raffinose, fructooligosaccharide, xylo-oligosaccharide, isomaltooligosaccharide, gentiooligosaccharide, or galactooligosaccharide. A bile acid adsorbing substance comprising an intestinal bacterium having a property of taking bile acid into a microbial cell in the presence of a sugar source. 7. The substance according to claim 6, wherein the intestinal bacterium is a Clostridium genus, a Collinsella genus, a Ruminococcus genus or a Bacteroides genus. 7. The substance according to claim 6, wherein the intestinal bacterium is a bacterium having no secondary bile acid producing activity. 7. The substance according to claim 6, wherein the intestinal bacterium is Clostridium butyricum. The substance according to claim 6, wherein the sugar source is glucose, raffinose, fructooligosaccharide, xylo-oligosaccharide, isomaltooligosaccharide, gentio-oligosaccharide, galactooligosaccharide. Food-drinks which have a bile acid adsorption effect containing the substance of any one of Claims 6-10. The bile acid adsorption agent which has a bile acid adsorption effect containing the substance of any one of Claims 6-10. A bile acid adsorbing substance having an action of adsorbing bile acids, characterized by comprising an assimilating sugar of an enteric bacterium having a property of taking bile acids into cells in the presence of a sugar source. 14. The substance according to claim 13, wherein the intestinal bacterium is a genus Clostridium, a Collinsella, a Ruminococcus, or a Bacteroides genus. 14. The substance according to claim 13, wherein the intestinal bacterium is a bacterium having no secondary bile acid producing activity. 14. The bile acid adsorbing substance according to claim 13, wherein the intestinal bacterium is Clostridium butyricum. 14. The substance according to claim 13, wherein the sugar source and the assimilating sugar are glucose, raffinose, fructooligosaccharide, xylo-oligosaccharide, isomaltoligosaccharide, gentio-oligosaccharide, and galactooligosaccharide. A food or drink having a bile acid adsorption action, comprising the substance according to any one of claims 13 to 17. A bile acid adsorbent comprising the substance according to any one of claims 13 to 17. A bile acid adsorbing substance having an action of adsorbing bile acids, comprising an intestinal bacterium having a property of taking bile acids into cells in the presence of a sugar source and its assimilating sugar. A method for adsorbing bile acids, comprising using an enteric bacterium having a property of taking bile acids into cells in the presence of a sugar source and an assimilating sugar thereof.