KR100663656B1 - An enzyme composition capable of forming an oligosaccharide - Google Patents

Abstract

In the present invention, oligosaccharides having physiological activity are synthesized in vivo, for example, to improve the intestinal flora. The present invention provides an enzyme composition comprising an enzyme capable of producing oligosaccharide having physiological activity in vivo and a method for producing in vivo a oligosaccharide having physiological activity.

Oligosaccharides, glucosyltransferases, fructosyltransferases, levansukrases, transglycosylation reactions, obesity, diabetes.

Description

An enzyme composition capable of forming an oligosaccharide

1 is a graph showing the results of Example 1 in relation to the effect of pH on the enzyme activity of glucosyltransferase.

FIG. 2 is a graph showing the results of Example 1 in relation to the effect of pH on the enzyme activity of Levansucrase. FIG.

3 is a graph showing the results of Example 1 in relation to the effect of pH on the enzyme activity of α-amylase.

4 is a graph showing the results of Example 2. FIG.

Figure 5 is a graph showing the results of Example 3, the resulting sugar composition is shown in the order of glucose, maltose, isomaltose, maltotriose, panos, lactosucrose and other sugars starting from below.

FIG. 6 is a graph showing the results of Example 7 wherein the sugars produced are shown as their total amounts in the order of other sugars and oligosaccharides starting from below.

FIG. 7 is a graph showing yet another result of Example 7, wherein the produced sugars are shown as their total amounts in the order of other sugars and oligosaccharides starting from below.

The present invention relates to an enzyme composition comprising an enzyme capable of producing an oligosaccharide having physiological activity in vivo, and also to a method for producing an oligosaccharide having physiological activity in vivo, including using the enzyme composition. .

Recently, with the progress of researches on the physiological action and function of carbohydrates, various useful sugars have been discovered, in particular, various oligosaccharides produced by glucosyltransferases, for example, fructooligosaccharides, soy oligosaccharides, galacto There is a high interest in oligosaccharides, xyloligosaccharides, genthiooligosaccharides, lactosucrose, coupling sugars and paratinose. Oligosaccharides are not only used as sweeteners, but also as sweeteners for diet by inhibiting digestion and absorption of foods as energy sources by their anti-affective or indigestible properties. In addition, since oligosaccharides are used as nutrients for lactobacteria and bifidobacterium and can regulate the intestinal environment, they are intended to improve the intestinal flora and improve physical conditions such as prevention of constipation or diarrhea and increased intestinal peristalsis. As it is used in specific health foods, their physiological activity is attracting attention.

Oligosaccharides have conventionally been produced in a variety of ways. That is, oligosaccharides are prepared on an industrial scale using various types of hydrolases, transferases and the like. As a method of supplying the oligosaccharides prepared as described above in vivo, the oligosaccharides are prepared by incorporating oligosaccharides as constituents of food ingredients such as carbonated drinks, soft drinks, household sugars, fermented milk, candy, biscuits, and chocolates. The method of making it show physiological activity or taking these oligosaccharides directly is performed.

However, when these various food ingredients are mixed with these oligosaccharides, the use of them is highly likely to suppress a number of problems, for example, the taste, flavor and texture of the food ingredients, and it is necessary to select an oligosaccharide suitable for each food ingredient. Problems occur. In other words, in order to actively ingest these oligosaccharides, it is necessary to selectively ingest the food ingredients containing the oligosaccharides, so that the diversity of dietary life cannot be satisfied.

An object of the present invention is to provide an enzyme composition comprising an enzyme capable of producing oligosaccharide having physiological activity in vivo and a method for producing oligosaccharide having physiological activity in vivo.

The inventors of the present invention have conducted various studies on a method for supplying oligosaccharides to a living body. As a result of extensive research on the possibility of producing oligosaccharides in vivo, the present inventors have found that the production of oligosaccharides in vivo by simultaneously ingesting enzymes capable of working in vivo with a food material containing a substrate for producing oligosaccharides. It has been found that this can be achieved to complete the present invention.

Accordingly, the present invention relates to an enzyme composition comprising an enzyme capable of producing oligosaccharides having physiological activity in vivo.

The present invention also provides the above-described enzyme composition in which the enzyme can act in the gastrointestinal tract, the enzyme composition in which the enzyme catalyzes the transglycosylation, the enzyme is glucosyltransferase, fructosyltransferase and levansukraze The enzyme composition is at least one enzyme selected from the group consisting of, the enzyme composition further comprises at least one enzyme selected from the group consisting of amylases and invertases and the enzyme composition whose physiological activity is a dietary effect.

The present invention also relates to a method for producing oligosaccharides having physiological activity in vivo, including using the enzyme composition. In addition, the present invention relates to a method in which the enzyme composition is ingested before meals, between meals or after a meal, and a method in which the enzyme composition is ingested with food.

As the enzyme capable of acting in vivo and synthesizing oligosaccharides in vivo by acting as a substrate on an ingredient taken as a food ingredient, any enzyme can be used as long as it can exhibit activity in an in vivo environment.

Examples of oligosaccharides having various physiological activities mentioned above include fructooligosaccharides (oligosaccharides in which 1-3 fructose molecules are bound to the fructose residues of sucrose at the C2 and C1 positions by β-bonding), branched oligosaccharides (oligosaccharides with α-1,6 bonds), galactooligosaccharides (such as raffinose and starchiose) and genthiooligosaccharides (oligosaccharides with β-1,6-glucoside bonds), and more specific examples thereof include 4-α-glucosyl-xylose, 3-α-glucosyl-sorbose, 4-α-glucosyl-sucrose, 4-α-glucosyl-mannose, 4-α-glucosyl-glucosamine, 4 -α-glucosyl-N-acetyl-glucosamine, 3-, 6-α-glucosyl-mannose, 3-, 4-α-glucosyl-xylose, 1-, 3-, 4-α-glucosyl Fructose, α-glucosyl-glycerol, riboflavin-α-glucoside, 6-α-galactosyl-fructose, 6-α-galactosyl-galactose, α-galactosyl-glycer Roll, 3-, 4-, 6-β-galactosyl-lactose, β-galactosyl-glycerol, 4-β-galactosyl-glucose, 4-β-galactosyl-mannose, β-galacto Cyl-glycerol, xyloxyl-fructoside, galactosyl-fructoside, isomaltosyl-fructoside, lactosyl-fructoside, 1-kestos, nitos (fructo-oligosaccharides), neokes Toss, inulobiose, difructofuranosyl 1,2 ': 2,1': 2,3 ': 2,6': 2 ', 6-2 anhydride and lactosurose.

These hetero-oligosaccharides can be synthesized using various types of glucosyltransferases. Examples of glucosyltransferases include those which act to transfer glucosyl groups, galactosyl groups and fructosyl groups. As enzymes that can be used to transfer glucosyl groups, transglucosidase, cyclodextrin synthetase, amylomalase, α-galactosidase and β-galactosidase are known and transfer of galactosyl groups Enzymes which can be used for the preparation are known as α-galactosidase, β-galactosidase and β-galactanase, and enzymes which can be used to transfer fructosyl groups include levansucrase, β- Fructofuranosidase and cyclic disaccharide synthetase are used.

For example, 4-α-glucosyl-xylose, 3-α-glucosyl-sorbose and 4-α-glucosyl-sucrose can be synthesized using cyclodextrin synthetase and 4-α- Glucosyl-mannose, 4-α-glucosyl-glucosamine and 4-α-glucosyl-N-acetyl-glucosamine can be synthesized using amylomalase, 3- and 6-α-glucosyl -Mannose, 3-, 4-α-glucosyl-xylose, 1-, 3-, 4-α-glucosyl-fructose, α-glucosyl-glycerol and riboflavin-α-glucoside are α-glucosidase 6-α-galactosyl-fructose, 6-α-galactosyl-galactose, α-galactosyl-glycerol, 3-, 4-, 6-β-galactosyl Lactose and β-galactosyl-glycerol can be synthesized using α-glucosidase, 4-β-galactosyl-glucose, 4-β-galactosyl-mannose and β-galactosyl- Glycerol can be synthesized using β-galactosidase , Xylosyl-fructoside, galactosyl-fructoside, isomaltosyl-fructoside and lactosyl-fructoside can be synthesized using levansucrase, 1-ketose, varnish Toss (fructo-oligosaccharides), neoketos, inulobiose, xylosyl-fructosides and galactosyl-fructosides can be synthesized using β-fructofuranosidase and difructofura Nosil 1,2 ': 2,1': 2,3 ': 2,6': 2 ', 6-2 anhydride can be synthesized using cyclic disaccharide synthetase.

These reactions can be expected to produce oligosaccharides by administering transferase alone if there are sufficient ingredients in the food material that can be used as substrates, but they will act on the food materials to produce ingredients that can be used as substrates for these transferases. It may be desirable and more effective to include other enzymes that can be used, such as amylases and invertases.

As the source of the enzymes mentioned above, any source can be useful, such as bacteria, fungi, yeast, actinomycetes, basidiomycetes, plants and animals, although the action of these enzymes is in vivo, in particular In the gastrointestinal, if necessary, inactivation in the low pH range of the gastrointestinal is a problem, so an enzyme should be selected that will have a sufficiently stable effect at low pH.

Examples of enzymes that can be used in the present invention are glucosyltransferases, which are enzymes that function to produce isomaltose, panos and other oligosaccharides by transferring glucose, and known sources thereof include microorganisms such as streptoco kusu (Streptococcus) genus Bacillus (Bacillus) genus Aspergillus (Aspergillus) genus Aureobasidium (Aureobasidium) in keulrep and when, for Ella (Klebsiella) in the plant, for example, include onion. More specifically, glucosyltransferase from Aspergillus niger (trade name: transglucosidase L "Amano", Amano Pharmaceutical Co., Ltd. (Amano Pharmaceutical Co., Ltd) .) Can be used.

Fructosyltransferases act primarily on sucrose to cleave β-1,2 bonds between fructose and glucose and then transfer the released fructose to sucrose to produce oligosaccharides, a known source of Silver microorganisms such as genus Bacillus, genus Arthrobacter , genus Aspergillus, genus Fusarium , genus Gloeosporium , genus Saccharomyces , genus Rhototorula ( Rhodotorula) genus Pichia (Pichia), a century Cronulla (Hansenula) and Candida genus (Candida), for in the plant, for example, include asparagus and Jerusalem Artichoke (Jerusalem artichoke). More specifically, the Bacillus natto (Bacillus natto) can be given a fructosyl transferase derived from the example [see: Denpun Kagaku, Vol. 38, no. 2, 217-222 (19991).

Levan sucrase is an enzyme that transfers the fructose residue of sucrose to produce various oligosaccharides such as high molecular weight polysaccharide levane, examples of which include levansukrases from Zymomonas mobilis IFO-13756 [see : Journal of Fermentation and Bioengineering, Vol. 79, No. 4, 367-369 (1995) and Rahnella aquatilis JCM-1683.

Amylase is an enzyme that acts on starch to produce disaccharides, trisaccharides, tetrasaccharides, and oligosaccharides, which are the substrates of glucosyltransferases. More specific examples thereof include amylases from Aspergillus oryzae . Biodiastase 2000, Amano Pharmaceutical Co., Ltd.].

In the present invention, these enzymes may be mixed with food ingredient components used as enzyme substrates, or may be administered to a living body separately or simultaneously with these food ingredients, but in order to obtain an effective action of these enzymes in vivo, It is important to establish a location where it can coexist with food ingredients that are enzyme substrates.

In general, food consumed from the oral cavity is digested through complex steps. In general, in the first step, the food is decomposed by α-amylase secreted from salivary glands, and in the second step, it is decomposed by pepsin under acidic conditions of hydrochloric acid in the stomach and finally secreted from the pancreas. It is broken down in the gut by enzymes and then absorbed into the intestinal wall as a nutrient.

As a result, when anticipating the effect of oligosaccharides having physiological activity, it is preferred that oligosaccharides be produced at an early stage of the digestive tract. That is, it is preferable that the oligosaccharide is produced by the reaction of the enzyme with various substrates in an acidic environment in the stomach.

After the food is ingested in vivo, in order for the enzyme composition of the present invention to be ingested at the same time in the gastrointestinal tract effectively, the enzyme constituting the enzyme composition must have sufficient properties to function under acidic conditions in the stomach. For this reason, among the above-mentioned enzymes, particularly preferred enzymes include glucosyltransferase derived from Aspergillus niger [trade name: transglucosidase L "Amano", Amano Pharmaceutical Co., Ltd., manufactured by Simonas Mobilis. Mention may be made of Levansukrases from IFO-13756 and Amylases from Aspergillus Oriza (trade name: Biodiastatase 2000, manufactured by Amano Pharmaceutical Co., Ltd.).

The dosage of the enzyme composition may be such that it exhibits an oligosaccharide-producing action in vivo, but may vary greatly depending on the nature and purity of each enzyme used, so the dosage should be set according to each property. For example, for the glucosyltransferases mentioned above, it can be 10,000 to 5,000,000 U / times, for Levansucrase, it can be 100 to 50,000 U / times, and for amylase, 10 to 5,000 U / times It may be one time.

In addition, in view of incompatibility issues, enzymes may optionally be prepared into enzyme compositions by mixing with other active ingredients. Of course, various auxiliaries can also be mixed for the preparation of the enzyme composition. In addition, agents that mix enzymes with antacids or act on the digestive tract, such as H ₂ blockers, can be used together.

As a method of administering the enzyme composition to a living body to act on a substrate as a food material, the enzyme composition can be used in a dosage form such as a powder, granule, solution, solid or capsule. In addition, the enzyme composition may be used in combination with various components for ingestion at the same time as the food material. Specifically, for example, the enzyme composition may be mixed with sucrose in a coating such as corn flakes and taken simultaneously with a meal.

The enzyme composition described above can usually be administered to a subject, preferably a mammal, more preferably a human, usually 1 to 3 times / day, such as a warm-blooded animal. The amount of oligosaccharides produced in vivo is about 2-5 g / day. Desired physiological activities of oligosaccharides include improving intestinal flora, suppressing constipation or diarrhea, promoting intestinal peristalsis and dieting (weight control such as obesity prevention).

Examples of the present invention are described below, but the present invention is not limited thereto.

In this regard, unless otherwise stated, each enzyme activity is measured by the following method.

Activity of Glucosyltransferases

Using α-methyl-D-glucoside as a substrate, the amount of enzyme that reacts the enzyme solution at 40 ° C., pH 5.0 and produces 1 μg of glucose for 60 minutes is defined as 1 U.

Activity of Levansukrases

It is measured using an F kit (D-glucose / D-fructose) (Boehringer-Mannheim GmbH). Using sucrose as a substrate, the amount of enzyme that produces 1 mg / ml of glucose in the reaction solution is defined as 1 U.

Amylase Activity

It is measured according to the method for testing starch glycation activity in the digestion activity test method described in Pharmacopoeia of Japan (General Test Methods) (37 ° C., pH 5). The amount of enzyme that increases the reducing power corresponding to 1 mg of glucose within 1 minute is defined as 1 U.

Example 1

Glucosyltransferase from Aspergillus Niger [trade name: transglucosidase L "Amano", Amano Pharmaceutical Co., Ltd., Levansukrase from Jimonas mobilis IFO-13756 [see: Journal of Fermentation and Bioengineering, Vol. 79, No. 4, 367-369 (1995)] and amylase from Aspergillus Oriza (trade name: Biodiastatase 2000, Amano Pharmaceutical Co., Ltd.) as the enzymes to be tested as pH for enzyme activity. Investigate the impact of

Test method (enzyme pH stability)

Each enzyme is treated at 37 ° C. for 1 hour in a buffer solution with each pH value to determine residual activity and the results are shown as relative to the highest activity defined as 100%. The results are shown in Figures 1-3.

Since all the enzymes tested exhibited sufficient residual activity at pH 3.5 to 6, it can be confirmed that these enzymes have properties that can be used for reaction in the stomach.

Example 2

Using the digestive tract model, the transfer reaction is carried out by subjecting levansukrase described in Example 1 to sucrose and lactose as substrates.

Substrate solution was added 6.86% sucrose, lactose 3.01%, gastric mucosa mucin 0.1%, Na ⁺ 150 mM and Ca ⁺ 1 mM in 13 mM acetate buffer (pH 4) containing 0.067 mg / ml pepsin, final pH 4.0 with hydrochloric acid It is prepared by adjusting.

Gastrointestinal Model: While maintaining the solution described above at 37 ° C., 500 U of Levansukrase was added slowly to react with stirring, and 0.1 N hydrochloric acid was added at intervals of 15 minutes until 70 minutes after the start of the reaction. It is used as a gastrointestinal environment until 120 minutes after the start of reaction. Sodium bicarbonate, sodium taurodeoxycholate and pancreatic enzyme are then added to use the reaction system as a gastrointestinal model. Some of the contents are sampled during the reaction and mixed with hydrochloric acid to terminate the reaction, followed by analysis by HPLC.

The results are shown in FIG. From this result, it can be confirmed that lactosucrose is produced under the gastrointestinal environment.

Example 3

Production of oligosaccharides by amylase, transglucosidase and levansukrases is measured using a gut model in the same manner as described in Example 2.

Substrate solution (150 mL) was dextrin (Pinedex: PD # 100), sucrose 6.86%, lactose 3.01%, gastric mucosa mucin 0.1%, Na ⁺ 150 mM, Ca ⁺ 1 mM and 13 mM acetate buffer (pH 4) It is adjusted to final pH 4.0 with hydrochloric acid.

Gastrointestinal model: While maintaining the substrate solution described above at 37 ° C., 10 mg pepsin, 70 mg biodiastatase, 100,000 U transglucosidase and 500 U lebansucrase were added and slowly stirred for reaction, until 70 minutes after the start of the reaction. 0.1N hydrochloric acid is added every 15 minutes. It is used as a gastrointestinal environment until 120 minutes after the start of reaction. Sodium bicarbonate, sodium taurodeoxycholate and pancreatic enzyme are then added to use the reaction system as a gastrointestinal model. Some of the contents are sampled during the reaction and mixed with hydrochloric acid to terminate the reaction, followed by analysis by HPLC.

The results are shown in FIG. From this result, it can be confirmed that various oligosaccharides such as isomaltose, panose and lactosucrose are produced under the gastrointestinal environment.

From this result, it can be clearly seen that 2 to 5 g / 1 day of oligosaccharide, which is considered an effective amount, can be sufficiently produced from the substrate in the food.

Example 4

When the method of Example 2 was repeated except that β-fructofuranosidase was used instead of levansucrase, lactosurose was produced.

Example 5

Seven healthy adults with low content of Bifidobacterium are administered the following enzyme composition for 14 days after eating. In this case, meals should be fed indefinitely without any special restrictions.

Amylase Part 5 Transglucosidase Part 5 Levansukraze Part 5 Filler (Lactose) Part 53 Binder (crystalline cellulose) Part 30 Glidants (Soft Silicate Anhydride) chapter 1 Lubricant (Magnesium Stearate) chapter 1

A mixture having the above composition is prepared into tablets according to the general method, and three tablets are administered after each meal.

Bifidobacterium cells in feces were measured and the results are shown in Table 1. Each data shown in Table 1 shows the logarithm of the average number of Bifidobacterium cells in 1 g of three times fecal samples of each adult. The results showed a significant increase in Bifidobacterium cell counts in five adult feces and enhanced bowel movement in all adults tested.

Before dosing 14 days after dosing One 8.8 9.7 ^* 2 7.5 8.8 ^* 3 8.2 9.0 ^* 4 9.1 9.2 5 8.3 8.9 ^* 6 8.5 8.4 7 7.8 9.3 ^{* *} Significant increase

In other words, when the enzyme composition is administered at mealtime, in vivo production of oligosaccharides is induced from various components contained in the food, oligosaccharides promote growth of Bifidobacterium, and are produced with the growth of Bifidobacterium. Acetic acid and lactic acid are thought to inhibit the growth of toxic bacteria in the intestine, thereby improving the intestinal flora and inhibiting the production of intestinal rot products to promote intestinal motility.

Example 6

Basic composition of the feed Mixed Feed (New Koromeal GS: Nippon Formula Feed Mfg.) 150 g dextrin 35 g Sucrose 10 g Lactose 5 g

Group 1: Feed consisting of basic composition and levansukrase (85 mg), biodiastatase (140 mg) and transglucosidase (310 mg)

Group 2: Feed and Levansukrase (85 mg) consisting of a basic composition

Third group: fodder consisting of basic composition

Pigs weighing about 7 kg are divided into 5 dogs per group for only 7 days of the basic composition, and then each of the above for 14 days. The weight gain rate per day and the number of cells of the intestinal Bifidobacterium were measured, and the results are shown in Tables 2 and 3. Each data shown in Table 3 represents the average logarithm of the number of Bifidobacterium cells in 1 g of enteric material of pigs belonging to each group.

Weight gain (g / day) First group 59.3 ± 16.6 Second group 59.4 ± 30.9 Third group 76.5 ± 15.5

Caecum Ascending Colon Descending colon rectal First group 7.9 8.4 8.8 8.1 Second group 7.3 7.5 7.8 7.8 Third group Less than 6.0 6.9 7.5 7.6

As is apparent from Tables 2 and 3, by using the enzyme composition of the present invention, unlike pigs reared in a general feed, the weight gain of the pigs is limitedly controlled, and the growth of intestinal bifidobacterium is also prevented. It is confirmed. In other words, it is thought that the enzymatic composition of the present invention produces indigestible oligosaccharides in vivo, resulting in such an effect of preventing obesity and an increase in useful enteric bacteria. From this result, when the diabetic patient uses the enzyme composition of the present invention, production and absorption of food ingredients that increase blood sugar can be suppressed, thereby suppressing blood sugar rise.

Example 7

Using corn flakes and milk, the oligosaccharide production test in the gastrointestinal model is performed as follows. To 40 g of Corn Flosty (trade name), milk (200 ml) is added and mixed, and the mixture is stirred at 37 ° C. for 30 minutes, and then the pH is adjusted to 4.5 ± 0.2. According to the method of Example 1, the enzyme composition of the following composition is added to and dissolved in 10 ml of the system described above.

(1) Levansukrase 50mg

(2) 50 mg Levansukrase

    Transglucosidase 183mg

    Biodiastatase-2000 250mg

After 10 and 30 minutes of reaction, samples were taken, centrifuged (3,000 rpm, 5 minutes) while cooling, and then 1 mL of the obtained supernatant was mixed with ₁ mL of 50 mM Na ₂ CO ₃ -NaH ₂ PO ₄ (pH 9.0). Mixing terminates the reaction. A 0.01 ml aliquot of the reaction terminated is mixed with 0.29 ml of water and 0.7 ml of acetonitrile, and the mixture is filtered and then analyzed by liquid chromatography. The results are shown in FIGS. 6 and 7. As can be clearly seen from FIGS. 6 and 7, oligosaccharides having physiological activity are produced after 10 and 30 minutes of reaction by using the enzyme composition of the present invention.

By the action of the enzyme composition provided by the present invention, oligosaccharides having physiological activity through food are produced from components as enzymatic substrates ingested in vivo, such that Intestinal flora can be improved. According to the present invention, oligosaccharides useful for a living body can be particularly used without considering their use, so that a diet effect and an effect of suppressing blood sugar increase in diabetic patients can be obtained.

While the present invention has been described in detail with reference to specific embodiments, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the spirit and scope of the invention.

This application is based on Japanese Patent Application No. 10-204293, filed July 2, 1998, and Japanese Patent Application No. 11-71122, filed March 17, 1999. Both Japanese patent applications are incorporated herein by reference.

According to the present invention, there is provided an enzyme composition comprising an enzyme capable of producing physiologically active oligosaccharides in vivo and a method of producing physiologically active oligosaccharides in vivo.

Claims (19)

Biologically active in vivo, comprising one or more enzymes selected from the group consisting of glucosyltransferases, fructosyltransferases and levansukrases, and additionally one or more enzymes selected from the group consisting of amylases and invertases An enzyme composition capable of forming an oligosaccharide selected from the group consisting of fructooligosaccharide, side chain oligosaccharide, galactooligosaccharide and genthio oligosaccharide. The enzyme composition of claim 1, wherein the enzyme can act in the stomach. delete delete delete The enzyme composition according to claim 1, wherein the physiological activity is an obesity prevention effect. The enzyme composition according to claim 1, wherein the physiological activity is a blood glucose raising inhibitory effect in a diabetic patient. delete delete delete delete delete delete delete The enzyme composition according to claim 1, which is ingested before meals, between meals or after meals. The enzyme composition according to claim 1, which is ingested with food. The enzyme composition according to claim 1 or 2 for promoting the growth of beneficial intestinal bacterial flora and for preparing a medicament for the prevention of constipation or diarrhea, intestinal peristalsis or weight control. Food comprising the enzyme composition according to claim 1. The enzyme composition according to claim 1 or 2, which is capable of forming an oligosaccharide selected from the group consisting of fructooligosaccharide, branched oligosaccharide, galactooligosaccharide and genthiooligosaccharide having physiological activity in vivo. .

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