Composition

Abstract

To provide a technique for further promoting various bacteria of the genus Bifidobacterium in intestinal flora.SOLUTION: A prebiotic composition for bacteria of the genus Bifidobacterium is obtained by combining 1-kestose and galactooligosaccharide. The composition can proliferate one or two or more types of bacteria of the genus Bifidobacterium selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition containing 1-kestose and a galactooligosaccharide. [Background technology]

[0002] It is known that the intestinal flora of infants is dominated by Bifidobacterium genus bacteria (hereinafter referred to as Bifidobacteria), and this is thought to be closely related to the maintenance of infant health. Therefore, developing foods that promote the growth of Bifidobacteria is important for maintaining the health of infants.

[0003] In recent years, there has been much research into prebiotics that promote the proliferation of bifidobacteria. Sugar sources in particular are considered to be important for the proliferation of bifidobacteria, and various oligosaccharides have been actively used. As one of these, 1-kestose has attracted attention as it promotes the proliferation of beneficial bacteria such as bifidobacteria and inhibits the proliferation of bacteria that are undesirable from the viewpoint of health (Non-Patent Document 1). Specifically, it has been reported that 1-kestose reduces Clostridium and promotes the proliferation of bifidobacteria even in an intestinal environment dominated by harmful bacteria in which the number of Clostridium is greater than the number of bifidobacteria (Patent Document 1). It has also been reported that 1-kestose inhibits the proliferation of bacteria that are undesirable from the viewpoint of health, such as bacteria of the genus Tizarella and Ruminococcus (Patent Documents 2 and 3).

[0004] In recent years, it has also been proposed to combine various oligosaccharides and use them as prebiotics. For example, it is known that a combination of fructooligosaccharides and galactooligosaccharides promotes the growth of bifidobacteria such as Bifidobacterium breve (Patent Documents 4 to 6). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2005-306781 A [Patent Document 2] JP 2020-097558 A [Patent Document 3] JP 2020-070245 A [Patent Document 4] International Publication No. 2005-39319 Brochure [Patent Document 5] International Publication No. 2005-110121 Brochure [Patent Document 6] International Publication No. 2006-91103 Brochure [Non-patent literature]

[0006] [Non-Patent Document 1] A. Endo et al., Anaerobe, 61 (2020) 102076. Summary of the Invention [Problem to be solved by the invention]

[0007] In order to effectively utilize the intake of prebiotics, it is necessary that the prebiotics are assimilated by intestinal bacteria and the intestinal bacteria grow. However, the genus Bifidobacterium includes many species, and the composition of the intestinal flora varies from individual to individual. In addition, there are various species of Bifidobacterium bacteria that are easily promoted in growth by prebiotics. Therefore, even if prebiotics are taken, some individuals may not have Bifidobacterium bacteria in their intestines that can assimilate the prebiotics, that is, they may not be "responders" to the prebiotics (=non-responders), and the effect of prebiotics may not be fully obtained. In view of such circumstances, an objective of the present invention is to provide a technique for further promoting various Bifidobacterium bacteria in the intestinal bacterial flora. [Means for solving the problem]

[0008] As a result of intensive research aimed at solving the above-mentioned problems, the inventors discovered that by combining 1-kestose and galactooligosaccharides, all four major species of Bifidobacterium bacteria can be grown evenly, and concluded that such a composition can be a prebiotic that is less likely to produce non-responders, thereby completing the present invention.

[0009] That is, a first aspect of the present invention is a composition containing 1-kestose and a galactooligosaccharide. In this embodiment, the mass ratio of the contents of 1-kestose and galactooligosaccharides is preferably 1:9 to 9:1. It is preferable that the composition of this embodiment further contains bacteria of the genus Bifidobacterium, including one or more species selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum. The composition of this embodiment is preferably a nutritional composition, more preferably a milk formula.

[0010] A second aspect of the present invention is a composition for promoting the growth of Bifidobacterium bacteria, which contains 1-kestose and galactooligosaccharides, wherein the Bifidobacterium bacteria include one or more species selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum.

[0011] A third aspect of the present invention is a composition for improving intestinal flora, which contains 1-kestose and galactooligosaccharides, wherein the intestinal flora includes Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum. Another aspect of this embodiment is a composition containing 1-kestose and galactooligosaccharides, which is administered or ingested by a subject suffering from a disease or condition that can be prevented or ameliorated by improving the intestinal flora, or a subject suffering from a disease or condition caused by deterioration of the intestinal flora, and which comprises Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum. The composition of this embodiment is preferably used for intestinal regulation, immune regulation, anti-allergy, defense against bacterial and viral infections, reduction of oxidative stress, prevention or amelioration of diarrhea, prevention or amelioration of constipation, prevention or amelioration of inflammatory bowel disease, or prevention of colon cancer.

[0012] In the second and third aspects, the mass ratio of the contents of 1-kestose and galactooligosaccharides is preferably 1:9 to 9:1. The compositions of the second and third aspects are preferably nutritional compositions, more preferably milk formulas. Effect of the Invention

[0013] According to the present invention, there is provided a composition capable of uniformly promoting the proliferation of various Bifidobacterium bacteria, particularly the four major species considered to be important for the health of infants, in the intestines. The composition of the present invention can be an excellent prebiotic that is unlikely to cause "non-responders" who are unlikely to obtain the proliferation effect of Bifidobacterium bacteria by ingestion. Such a composition can be in the form of an oral composition for foods, beverages, medicines, etc., or can be contained in foods, beverages, medicines, etc. as an additive, etc. Ingestion of the composition of the present invention is expected to provide an effect of improving the intestinal flora, and to be useful in maintaining the health of the person who consumes it, particularly infants and young children. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present invention will now be described in detail. However, the present invention is not limited to the following embodiments and can be freely modified within the scope of the present invention.

[0015] The composition of the present invention essentially contains 1-kestose and galactooligosaccharides.

[0016] 1-kestose is a trisaccharide (1-O-(β-D-fructofuranosyl)-β-D-fructofuranosyl-α-D-glucopyranoside, Glc α1→2 Fru β1→2 Fru) formed by binding one glucose and two fructose. 1-kestose can be produced by reacting various enzymes with sucrose using a known method, for example, the method described in JP-A-58-201980. High-purity 1-kestose can be obtained by purifying it using the chromatographic method described in JP-A-2000-232878 and then crystallizing it using the method described in JP-B-6-70075. Commercially available 1-kestose (for example, manufactured by Bussan Food Science Co., Ltd.) can also be used. The purity of 1-kestose in the crystals or raw material is preferably 95% or more, but may be 80% or more, or may be 70% or more. In the composition of the present invention, the content of 1-kestose is preferably 0.1 to 20 mass % based on the total composition (solid content), more preferably 0.2 to 5 mass %, and even more preferably 0.3 to 3 mass %. In addition, from the viewpoint of obtaining the desired effect of promoting the growth of Bifidobacterium bacteria, the composition of the present invention does not need to contain other fructooligosaccharides, for example, fructooligosaccharides with longer sugar chains such as nystose (tetrasaccharide) and 1-fructofuranosyl-D-nystose (pentasaccharide).

[0017] Galactooligosaccharides (GOS) are oligosaccharides or mixtures thereof having a structure represented by Gal-(Gal)n-Glc (n is 1-3, β1→4 bond or β1→6 bond). Galactooligosaccharides are industrially produced from lactose as a raw material by a transfer reaction using β-galactosidase, and the main component is 4'-galactosyllactose (4'-GL), a trisaccharide in which one galactose is bound to the non-reducing end of lactose. Commercially available galactooligosaccharides (e.g., Yakult Pharmaceutical Co., Ltd.) can also be used. Galactooligosaccharides may be one type or a mixture of two or more types. In the composition of the present invention, the content of the galactooligosaccharide is preferably 0.1 to 20 mass % based on the total composition (solid content), more preferably 0.2 to 5 mass %, and even more preferably 0.3 to 3 mass %.

[0018] As long as the composition of the present invention contains 1-kestose and galactooligosaccharides, it may contain one or more other oligosaccharides, such as lactulose, raffinose, fructooligosaccharides, soybean oligosaccharides, lactose oligosaccharides, xylooligosaccharides, isomaltulose, human milk oligosaccharides, coffee bean mannooligosaccharides, gluconic acid, polydextrose, and inulin.

[0019] In the composition of the present invention, the mass ratio of the contents of 1-kestose and galactooligosaccharides is preferably 1:9 to 9:1, more preferably 3:7 to 7:3, and even more preferably 4:6 to 6:4.

[0020] The composition of the present invention promotes the growth of bacteria belonging to the genus Bifidobacterium. It is possible to do so. The Bifidobacterium bacteria whose growth is promoted by the composition of the present invention include one or more species selected from Bifidobacterium longum (reclassified as Bifidobacterium longum subsp. longum), Bifidobacterium breve, Bifidobacterium infantis (reclassified as Bifidobacterium longum subsp. infantis), and Bifidobacterium bifidum. These four species of Bifidobacteria are particularly important for the health of the intestinal flora during infancy (hereinafter, also referred to as "infant-type Bifidobacteria"), and the composition of the present invention can promote the growth of these species evenly. In addition, as long as the growth of one or more of the above four species is promoted, the growth of other Bifidobacterium bacteria may also be promoted, such as Bifidobacterium adolescentis, Bifidobacterium catenulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium pseudolongum, and Bifidobacterium longum subsp. suis.

[0021] In this specification, "growth" of bacteria includes an increase in the absolute number of bacteria, and "promotion of growth" refers to a degree of increase that is greater when the composition of the present invention is applied than when it is not applied. That is, when the composition of the present invention is applied in vivo or in vitro, the effect of increasing the number of bacteria of the genus Bifidobacterium compared to when it is not applied is obtained. The degree of increase in the number of bacteria is not particularly limited, but it means that the number of bacteria is preferably 1.1 times or more, more preferably 1.5 times or more, and even more preferably 3 times or more, compared to the number of bacteria when the composition of the present invention is not applied. Such an increase in the number of bacteria can be confirmed not only by directly measuring the number of bacteria, but also by, for example, measuring the turbidity (absorbance) or the amount of short-chain fatty acids such as acetic acid in the digestive tract contents of a culture medium in which the bacteria have been cultured or of an animal such as a human that has ingested the composition, and observing an increase in these values, or by measuring the pH in the culture medium and observing a decrease in this value, etc.

[0022] Furthermore, "growth" includes an increase in the proportion of the bacteria in the intestinal flora, and "promotion of growth" refers to a greater degree of increase when the composition of the present invention is applied than when it is not applied. That is, it includes increasing the proportion of Bifidobacterium bacteria present in the digestive tract of an animal such as a human that has ingested the composition of the present invention. Here, "presence" can also be rephrased as "occupancy rate" of the entire group of bacteria detected in the intestinal flora. "Increase in presence" may mean an increase or decrease in the presence of other bacteria in the intestinal flora, as long as the presence of Bifidobacterium bacteria in the intestinal flora increases. The degree of such an increase in the presence ratio is not particularly limited, but is preferably at least 2% higher, more preferably at least 5%, and even more preferably at least 20% higher than the presence ratio of the bacteria when the composition of the present invention is not applied.

[0023] Furthermore, "promotion of growth" may refer to an increase in the rate at which the absolute number of bacteria increases, or the rate at which the proportion of the bacteria in the intestinal flora increases. The degree of increase in the rate is not particularly limited, but is preferably 10% or more, more preferably 20% or more, and even more preferably 50% or more higher than the rate at which the bacteria increases when the composition of the present invention is not applied. The increase in speed can be achieved, for example, by increasing the time required to reach a given number or proportion of bacteria using the present invention. This can be confirmed by, for example, determining whether the time is shorter when the composition is applied than when the composition is not applied.

[0024] The composition of the present invention may be in the form of a food, drink, medicine, etc. by itself, or may be in the form of being contained in a food, drink, medicine, etc. as an additive. The composition of the present invention may be administered orally or parenterally, but is usually administered orally. Parenteral administration includes rectal administration.

[0025] As described above, the composition of the present invention can promote the growth of Bifidobacterium bacteria, and therefore it is preferable for the composition to contain Bifidobacterium bacteria together with 1-kestose and galactooligosaccharides. Here, the Bifidobacterium bacteria contained in the composition of the present invention are not particularly limited, but include Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium catenulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium pseudolongum, etc. Among these, it is more preferable to contain one or more species selected from Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum.

[0026] Bifidobacterium longum is classified as Bifidobacterium longum NITE BP-02621 (also known as BB536 or Bifidobacterium longum subsp. longum ATCC BAA-999) can be used. Bifidobacterium longum BB536 was deposited in the NPMD under the NITE accession number NITE BP-02621 on January 26, 2018, based on the Budapest Treaty. The same bacterium, Bifidobacterium longum subsp. longum ATCC BAA-999 (number: ATCC BAA-999), is available from the American Type Culture Collection (ATCC: 10801 University Boulevard, Manassas, VA 20110, United States of America) as ATCC BAA-999 (see, for example, JP 2012-223134 A). In addition, as Bifidobacterium longum, Bifidobacterium longum subsp. longum ATCC 15707 available from ATCC can be used.

[0027] Examples of Bifidobacterium breve include Bifidobacterium breve M-16V. Bifidobacterium breve M-16V was deposited internationally under the Budapest Treaty at the National Institute of Technology and Evaluation (NPMD) (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818) on January 26, 2018, under the accession number NITE BP-02622. Commercially available products, such as "Bifidobacterium breve M-16V" manufactured by Morinaga Milk Industry Co., Ltd., may also be used. Another example of Bifidobacterium breve is Bifidobacterium breve MCC1274. Bifidobacterium breve MCC1274 has been internationally deposited on August 25, 2009, under the Budapest Treaty at the National Institute of Advanced Industrial Science and Technology (currently the National Institute of Technology and Evaluation Patent Organism Depositary (IPOD) (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818) under the deposit number FERM BP-11175.

[0028] Bifidobacterium infantis also includes Bifidobacterium infantis M-63, which was internationally deposited in the NPMD on January 26, 2018 under the Budapest Treaty under the accession number NITE BP-02623. Furthermore, as Bifidobacterium infantis, Bifidobacterium longum subsp. infantis ATCC 15697 available from ATCC can be used.

[0029] An example of Bifidobacterium bifidum is Bifidobacterium bifidum MCC 1092. Bifidobacterium bifidum MCC 1092 was internationally deposited with the NPMD on February 21, 2017 under the accession number NITE BP-02429 based on the Budapest Treaty. In addition, an example of Bifidobacterium bifidum is Bifidobacterium bifidum MCC 1319. Bifidobacterium bifidum MCC 1319 was internationally deposited with NPMD on February 21, 2017 under the accession number NITE BP-02431 based on the Budapest Treaty. In addition, an example of Bifidobacterium bifidum is Bifidobacterium bifidum MCC 1868. Bifidobacterium bifidum MCC 1868 was internationally deposited with NPMD on February 21, 2017 under the accession number NITE BP-02432 based on the Budapest Treaty. Furthermore, an example of Bifidobacterium bifidum is Bifidobacterium bifidum MCC 1870. Bifidobacterium bifidum MCC 1870 was internationally deposited with NPMD on February 21, 2017 under the accession number NITE BP-02433 based on the Budapest Treaty.

[0030] The bacteria specified by the above-mentioned examples of bacterial names are not limited to the strains deposited or registered in a given institution under the bacterial name (hereinafter, for convenience of explanation, also referred to as "deposited strains"), but also include strains substantially equivalent thereto (also referred to as "derived strains" or "derived strains"). In other words, they are not limited to the strains deposited in the depository institution under the above-mentioned accession numbers, but also include strains substantially equivalent thereto. For each bacterium, "strains substantially equivalent to the above-mentioned deposited strains" refers to strains that belong to the same species as the above-mentioned deposited strains, have an effect of improving the intestinal flora, and further have an identity of preferably 99.86% or more, more preferably 99.93% or more, and even more preferably 100% to the base sequence of the 16SrRNA gene of the above-mentioned deposited strain, and preferably have the same bacteriological properties as the above-mentioned deposited strain. For each bacterium, the strain substantially equivalent to the above-mentioned deposited strain may be, for example, a derived strain with the above-mentioned deposited strain as a parent strain. Derivative strains include strains bred from the deposited strain and strains that have arisen naturally from the deposited strain. Breeding methods include modification by genetic engineering techniques and modification by mutation treatment. Mutation treatments include irradiation with X-rays, irradiation with ultraviolet light, and treatment with mutagens such as N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, and methyl methanesulfonate. Strains that have arisen naturally from the deposited strain include strains that have arisen naturally during use of the deposited strain. Such strains include mutant strains that have arisen naturally by culturing (e.g., subculturing) the deposited strain. Derivative strains may be constructed by one type of modification, or by two or more types of modifications.

[0031] As the Bifidobacterium bacteria to be contained in the composition of the present invention, a commercially available product may be used, or one obtained by appropriate production may be used. Furthermore, the cells of the Bifidobacterium bacteria contained in the composition of the present invention can be easily obtained by culturing the above-mentioned Bifidobacterium bacteria. The culturing method is not particularly limited as long as the Bifidobacterium bacteria can grow. The culturing method can be any method that can be used. For example, a method commonly used for culturing Bifidobacterium bacteria can be used as is or with appropriate modifications. The culture temperature may be, for example, 25 to 50°C, and preferably 35 to 42°C. The culture can be preferably carried out under anaerobic conditions, for example, while aerating anaerobic gas such as carbon dioxide gas. The culture can also be carried out under microaerobic conditions such as liquid static culture. The culture can be carried out, for example, until the Bifidobacterium bacteria grow to a desired extent.

[0032] The medium used for the culture is not particularly limited as long as the Bifidobacterium bacteria can grow. As the medium, for example, a medium usually used for culturing Bifidobacterium bacteria can be used as it is or after appropriate modification. That is, as the carbon source, for example, sugars such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch, starch hydrolysate, blackstrap molasses, etc. can be used depending on the assimilation. As the nitrogen source, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, ammonium nitrate, and nitrates can be used. In addition, as the inorganic salt, for example, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, ferrous sulfate, etc. can be used. In addition, organic components such as peptone, soybean flour, defatted soybean meal, meat extract, yeast extract, etc. may be used. Specific examples of media commonly used for culturing Bifidobacterium include Reinforced Clostridial medium, de Man, Rogosa, and Sharpe medium (MRS medium), modified MRS medium (mMRS medium), TOS propionate medium (TOSP medium), and TOS propionate mupirocin medium (TOSP Mup medium).

[0033] As the Bifidobacterium bacteria that can be contained in the composition of the present invention, the bacterial cells or a fraction containing the bacteria can be used without any particular limitation. That is, as the Bifidobacterium bacteria, for example, a culture obtained by culturing may be used as it is, a diluted or concentrated culture may be used, or bacterial cells recovered from the culture may be used. In addition, various additional operations such as heating and freeze-drying may be performed after the culture as long as the effect of improving the intestinal flora is not impaired. The additional operation is preferably one that has high survival rate of the bacterial cells. That is, specific examples of the Bifidobacterium bacteria that can be contained in the composition of the present invention include a culture of Bifidobacterium bacteria, bacterial cells recovered from the culture, and processed products thereof, and the processed products include a diluted product, a concentrated product, or a dried product. It is preferable that the bacterial cells are usually used in a form containing live bacterial cells. The bacterial cells may be, for example, a product consisting of live bacterial cells, or a mixture of live bacterial cells and dead bacterial cells.

[0034] In the present invention, the Bifidobacterium bacteria contained in the composition include at least live bacteria, and the total amount of the bacteria in the composition is preferably 1 × 10 4 ~1×10 13 cfu / g or 1×10 4 ~1×10 13 cfu / mL, more preferably 1×10 5 ~1×10 12 cfu / g or 1×10 5 ~1×10 12 cfu / mL, more preferably 1×10 6 ~1×10 11 cfu / g or 1×10 6 ~1×10 11 It contains live Bifidobacterium bacteria at a concentration of cfu / mL. As long as it contains live bacteria, it may also contain dead bacteria. In addition, cfu refers to colony forming unit. In the present specification, for example, the value can be the value when cultured at 38° C. in a solid medium containing 10% by mass of reconstituted skim milk powder. These ranges may be the contents in the composition when it is normally distributed, or the contents when it is orally ingested.

[0035] As described above, the composition of the present invention can promote the growth of Bifidobacterium bacteria, including infantile bifidobacteria, and is therefore useful for promoting the growth of Bifidobacterium bacteria. This can be preferably applied.

[0036] Here, the Bifidobacterium bacteria to be grown are not limited to the Bifidobacterium bacteria contained in the composition as described above, but also include Bifidobacterium bacteria present in the digestive tract of animals such as humans that have been orally ingested. The composition of the present invention is used to grow preferably one or more species, more preferably all four species, of the four types of infantile bifidobacteria. Therefore, the composition of the present invention can be used to improve the intestinal flora. Here, "improving the bacterial flora" includes increasing the number of bacteria present in the intestinal flora of Bifidobacterium bacteria, particularly infantile Bifidobacteria, and the proportion of the bacteria present therein. In addition, "improving the bacterial flora" may include increasing the proportion of other good bacteria present in the intestinal flora or decreasing the proportion of bad bacteria present in the intestinal flora, so long as the proportion of Bifidobacterium bacteria present in the intestinal flora increases. Examples of other good bacteria include lactic acid bacteria. Examples of bad bacteria include Clostridium perfringens, Salmonella, Staphylococcus aureus, and pathogenic Escherichia coli. The "abundance ratio" can also be rephrased as the "occupancy rate" of the entire bacterial group detected in the intestinal bacterial flora.

[0037] The composition of the present invention can be useful for subjects with diseases or conditions that can be prevented or improved by improving the intestinal flora, or for subjects with diseases or conditions caused by the deterioration of the intestinal flora, for example, for intestinal regulation, immunoregulation, antiallergy, defense against bacterial and viral infections, reduction of oxidative stress, prevention and improvement of diarrhea, prevention and improvement of constipation, inflammatory bowel disease, prevention of colon cancer, etc.

[0038] The present invention can be rephrased as the use of 1-kestose and galactooligosaccharides in the production of a composition for promoting the growth of Bifidobacterium bacteria, wherein the Bifidobacterium bacteria include one or more species selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum. The present invention can be rephrased as the use of 1-kestose and galactooligosaccharides in promoting the growth of Bifidobacterium bacteria, wherein the Bifidobacterium bacteria include one or more species selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum. The present invention can also be said to be a composition containing 1-kestose and galactooligosaccharides used to promote the growth of Bifidobacterium bacteria, wherein the Bifidobacterium bacteria comprises one or more species selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum.

[0039] The present invention can also be said to be a method for promoting the growth of Bifidobacterium bacteria, which comprises administering 1-kestose and galactooligosaccharides to a subject, wherein the Bifidobacterium bacteria comprises one or more species selected from the group consisting of Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum. Here, the subject is not particularly limited as long as it is an animal, but is usually a human. Note that "administering to a subject" may be synonymous with "allowing a subject to ingest." Ingestion may be voluntary (free intake) or forced (forced intake). That is, the administration step specifically includes, for example, blending 1-kestose and galactooligosaccharides into food, drink, or feed and supplying them to the subject, thereby allowing the subject to freely ingest 1-kestose and galactooligosaccharides. The process may be a process of causing the

[0040] The timing of intake (administration) of the composition of the present invention is not particularly limited, and can be appropriately selected depending on the condition of the subject to be administered.

[0041] The intake (administration) amount of the composition of the present invention is appropriately selected depending on the age, sex, condition, other conditions, etc. of the subject to be ingested (administered). Regardless of the amount or duration of intake (administration), the drug can be administered once a day or in multiple divided doses.

[0042] The composition of the present invention is preferably in the form of a food or drink. There are no particular limitations on the form or properties of the food or drink as long as the effects of the present invention are not impaired, and the food or drink can be produced by ordinary methods using ingredients normally used for food or drink. The composition of the present invention also includes additives to be added to foods, beverages, and medicines. Examples of such additives include additives to be added to expressed breast milk or formula milk, and it is assumed that the milk after the additives are administered to newborns or infants. Food and drink are usually taken orally, but are not limited thereto, and may be taken, for example, nasally, or through a gastrostomy or enterostomy. For example, it is envisaged that newborns and infants will be given formula, which is the composition of the present invention described below, or breast milk to which the composition of the present invention has been added, through a nasogastric feeding tube or the like.

[0043] Food and drink products include, regardless of their form, liquid, paste, gel-like solid, powder, etc., for example, tablet confectionery; flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken flour, breadcrumbs, etc.; instant noodles, cup noodles, retort / prepared foods, pre-cooked canned foods, microwave foods, instant soups / stews, instant miso soup / cleansing liquids, canned soups, freeze-dried foods, other instant foods, etc.; canned agricultural products, canned fruit, jams / marmalades, pickles, Agricultural processed products such as boiled beans, dried agricultural goods, cereals (processed grain products); marine processed products such as canned seafood, fish ham and sausage, seafood paste products, seafood delicacies, and tsukudani (fried fish stew); livestock processed products such as canned livestock and paste, livestock ham and sausage; dairy products such as processed milk, milk drinks, yogurt, lactic acid bacteria drinks, cheese, ice cream, cream, and other dairy products; fats and oils such as butter, margarines, vegetable oils, soy sauce, miso, sauces, processed tomato seasonings, mirin , vinegars, and other basic seasonings; complex seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other complex seasonings; frozen foods such as frozen ingredients, semi-cooked frozen foods, and cooked frozen foods; confectioneries such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice snacks, bean snacks, dessert snacks, jellies, and other sweets; beverages such as carbonated drinks, natural fruit juice, fruit juice drinks, soft drinks with fruit juice, fruit pulp drinks, fruit drinks with fruit particles, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic drinks, and other beverages; other commercially available foods such as baby food, furikake, and ochazuke nori seaweed; nutritional compositions such as prepared milk (including powdered milk, liquid milk, etc.), liquid foods, and supplements; health functional foods (foods for specified health uses, foods with nutritional functions, and foods with functional claims).

[0044] Of these, nutritional compositions are preferred. In the present invention, the "nutritional composition" is an embodiment of a food or drink, and is not particularly limited, but is preferably a formula, a liquid diet, a supplement, etc., and more preferably a formula. The subjects of intake may be infants, small children, children, or adults, but are preferably infants and small children. Formulated milk includes powdered milk and liquid milk. In the Ministerial Ordinance on the Ingredient Standards of Milk and Dairy Products (Milk Ministerial Ordinance), modified milk powder is defined as "a product that is made by processing raw milk, cow's milk, special cow's milk, or foods made using these as ingredients, or that uses them as the main ingredient." It is defined as "a powder made by adding nutrients necessary for infants and young children." The ministerial ordinance defines modified liquid milk as "a liquid made by processing raw milk, cow's milk, special cow's milk, or foods made using these as ingredients, or using them as the main ingredient, and adding nutrients necessary for infants." In addition, the modified milk is a mixture of various nutritional components such as proteins, fats and oils, carbohydrates, minerals, and vitamins, and includes those processed into powder or liquid form. In addition, prepared milk further includes "powdered milk formula for infants," "liquid milk formula for infants," and "powdered milk formula for pregnant and lactating women," which are foods for special dietary uses defined in the Health Promotion Act, as well as forms such as powdered milk formula for young children, nutritional powder for adults, and nutritional powder for the elderly.

[0045] When the composition of the present invention is in the form of a supplement, it can be formulated into solid preparations such as powders, granules, tablets, capsules, etc.; liquid preparations such as solutions, syrups, suspensions, emulsions, etc. When formulating such preparations, the components, carriers, and methods for formulating pharmaceuticals described below can be followed.

[0046] In addition, one aspect of the food and drink may be feed, such as pet food, livestock feed, and fish feed. The form of the feed is not particularly limited, and may contain, for example, grains such as corn, wheat, barley, rye, milo, etc.; vegetable oil cakes such as soybean oil cake, rapeseed oil cake, palm oil cake, linseed oil cake, etc.; bran such as wheat bran, wheat bran, rice bran, defatted rice bran, etc.; manufacturing residues such as corn gluten meal and corn jam meal; animal feeds such as fish meal, skim milk powder, whey, yellow grease, tallow, etc.; yeasts such as torula yeast and brewer's yeast; mineral feeds such as calcium phosphate and calcium carbonate; oils and fats; simple amino acids; sugars, etc.

[0047] When the composition of the present invention is in the form of a food or drink (including feed), it can be provided and sold as a food or drink labeled for its use in proliferation of bifidobacteria in the intestine.

[0048] Such "indication" acts include all acts aimed at informing consumers of the aforementioned uses, and any expression that can recall or infer the aforementioned uses falls under the category of "indication" acts of the present invention, regardless of the purpose of the indication, the content of the indication, the object or medium on which it is displayed, etc. Furthermore, it is preferable that the "labeling" be done in an expression that allows consumers to directly recognize the above-mentioned uses.Specific examples include the act of transferring, delivering, displaying for the purpose of transferring or delivering, or importing food and beverage products or product packaging on which the above-mentioned uses are written, displaying or distributing advertisements, price lists, or transaction documents related to the products and writing the above-mentioned uses in information containing the above-mentioned uses and providing them by electromagnetic means (such as the Internet), etc.

[0049] On the other hand, the content of the labeling is preferably a labeling approved by the government etc. (for example, a labeling approved based on various systems established by the government and made in a manner based on such approval, etc.) In addition, it is preferable that such a labeling content is affixed to the packaging, containers, catalogs, pamphlets, POP and other sales site promotional materials, other documents, etc.

[0050] Furthermore, "labeling" also includes labeling as health food, functional food, enteral nutrition food, special purpose food, health functional food, food for specified health uses, nutrient functional food, functional food, medical drug, etc. Among these, in particular, labeling approved by the Consumer Affairs Agency, such as labeling approved under systems related to foods for specified health uses, nutrient functional foods, or foods with functional claims, or systems similar to these, can be mentioned. Specifically, labeling as a food for specified health uses, labeling as a food for conditional specified health uses, labeling that indicates an effect on the structure or function of the body, labeling that reduces the risk of disease, labeling of functionality based on scientific evidence, etc. can be mentioned, and more specifically, labeling approved under the Cabinet Office Ordinance on the Permission of Labeling for Special Purposes Provided in the Health Promotion Act (effective as of August 31, 2009) can be mentioned. Typical examples are labeling as a food for specified health uses (particularly labeling of health uses) as stipulated in Cabinet Office Ordinance No. 57 and similar labeling. Examples of such claims include "for those who want to increase the number of bifidobacteria," "to increase the number of bifidobacteria that are beneficial to babies' health," "to improve intestinal flora," "for the stomach health of infants and young children," etc.

[0051] When the composition of the present invention is in the form of a pharmaceutical product, the route of administration may be either oral or parenteral, but oral is preferred. Parenteral ingestion (administration) includes rectal administration, etc. The pharmaceutical form can be formulated into a desired dosage form depending on the administration method. For example, in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets, and capsules; liquid preparations such as solutions, syrups, suspensions, and emulsions. In addition, in the case of parenteral administration, it can be formulated into suppositories, ointments, injections, and the like. In the formulation, ingredients such as excipients, pH adjusters, colorants, flavorings, etc. that are usually used in formulations can be used. In addition, it is also possible to use other medicinal ingredients, prebiotics against known or future Bifidobacterium bacteria, prebiotics against other bacteria, etc. in combination. In addition, formulation can be carried out by a known method as appropriate depending on the dosage form. When preparing the formulation, a pharmaceutical carrier may be appropriately added to the formulation.

[0052] Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose, and calcium carboxymethyl cellulose; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

[0053] Examples of binders include gelatin, polyvinylpyrrolidone, macrogol, and the like, in addition to the above-mentioned excipients.

[0054] Examples of disintegrants include the above-mentioned excipients, as well as chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone.

[0055] Examples of lubricants include talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as veegum and glomerulus; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylates such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; starch derivatives, and the like.

[0056] Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

[0057] Examples of flavoring agents include sweeteners, acidulants, and fragrances. In the case of a liquid preparation for oral administration, examples of the carrier used include solvents such as water.

[0058] The timing of taking the pharmaceutical agent of the present invention is not particularly limited, and may be before or after a meal, between meals, or before going to bed. EXAMPLES

[0059] The present invention will be described in more detail below using examples, but the present invention is not limited to these examples.

[0060] <Test Example 1> Evaluation of the growth-promoting effect of various oligosaccharides on infant bifidobacteria by fecal culture (1) Sample pretreatment Stool samples were obtained from seven healthy infants aged 1 to 12 months (referred to as a, b, c, d, e, f, and g). They were collected immediately after defecation and immediately transferred to anaerobic conditions at 10°C or less. Within 8 hours after defecation, the collected stool was diluted with 10 times the volume of saline and stored at -80°C until use.

[0061] (2) Culture test In order to simulate the environment in the large intestine, a culture experiment was carried out according to the following procedure. A Bio Jr.8 100 mL x 8-connected culture device (BJR-25NA1S-8M manufactured by Biot Co., Ltd.) was used, and the operation was performed according to the instruction manual for the device. 100 mL of YCFA medium with the composition shown in Table 1 was used as the medium. The total concentration of oligosaccharides in the medium was 1 mass%. The medium components other than the oligosaccharides were dissolved in purified water and sterilized by autoclave, and the oligosaccharides were dissolved in purified water, sterilized by filtration, and added to the autoclaved mixture. The culture temperature was 37°C, and filter-sterilized CO was added to each culture vessel. 2The culture was maintained in an anaerobic state during the incubation period. After adjusting the pH to 7.0, 100 μL (10 mg of stool) of a stool sample diluted with saline was added, and anaerobic incubation was started. In order to mimic the pH in the infant's large intestine, 1 M Na 2 CO 3 The medium was collected 24 hours after the start of culture and centrifuged at 8000 rpm for 3 minutes at 4°C, and the supernatant and precipitate were collected.

[0062] [Table 1]

[0063] (3)DNA extraction DNA was extracted from the precipitate collected in (2) using a GENE PREP STAR PI-480 automatic extractor (Kurabo Industries Ltd Japan). Using the extracted DNA as a template, the number of Bifidobacterium bacteria and various Bifidobacterium species (Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium infantis, and Bifidobacterium bifidum) was measured by quantitative PCR using an Applied Bio systems 7500 Fast & 7500 real-time PCR system (Thermo Fisher). The primers and PCR conditions are shown in Table 2. Note that whether Table 2-1 or 2-2 is used, the results are the same. good.

[0064] [Table 2-1]

[0065] [Table 2-2]

[0066] (4) Results Tables 3 to 7 show the growth rates of the genus Bifidobacterium and various species of Bifidobacterium after 24 hours of culture, measured by quantitative PCR. The growth rate is a logarithmic value of the increase rate when the number of bacteria before culture is set to 1. The species and number of Bifidobacterium bacteria possessed by the test infants ranged from 1 to 4 species, consistent with previous reports, and the combinations were also diverse for each individual. The growth promotion effect on the genus Bifidobacterium and each species of Bifidobacterium was analyzed by analysis of variance and analysis of covariance ("Fit to Model" menu of statistical analysis software JMP), and each group (Example / Comparative Example) was ranked in order of the growth effect, with the top group placed in the upper row of the table. In addition, each sample was ranked in order of the growth effect, with the top sample placed in the left column of the table. In other words, each group and each sample was ranked by comprehensive evaluation using statistical processing, and arranged from the top left to the bottom right in order of decreasing growth effect.

[0067] Galactooligosaccharides alone showed excellent growth-promoting effects on Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum, but did not show sufficient growth-promoting effects on Bifidobacterium longum (Comparative Example 1). On the other hand, when 1-kestose and galactooligosaccharides were combined, the excellent growth-promoting effects on Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum were not lost, and Furthermore, it also showed an excellent growth-promoting effect on Bifidobacterium longum, and was ranked first or second among various combinations of oligosaccharides for the growth-promoting effect on all four types of infantile-type bifidobacteria (Example 1). This demonstrated that the combination of 1-kestose and galactooligosaccharides can uniformly promote the growth of all infantile-type bifidobacteria in all test infants. In other words, the combination of 1-kestose and galactooligosaccharides can be an excellent prebiotic that is unlikely to cause non-responders.

[0068] [Table 3]

[0069]

Table 4

[0070]

Table 5

[0071]

Table 6

[0072]

Table 7

Claims

[Claim 1] A composition containing 1-kestose and a galactooligosaccharide.

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