Nutritional composition

Abstract

To provide a technique for further promoting the growth of Bifidobacterium bacteria in intestinal bacterial flora.SOLUTION: Provided is a nutritional composition containing an oligosaccharide and a tocopherol and in which the tocopherol is one or more selected from the group consisting of β-tocopherol, γ-tocopherol and δ-tocopherol (however, excluding breast milk). The oligosaccharide preferably is human milk oligosaccharide. In the nutritional composition, the mass ratio (δ / β) of the δ-tocopherol content to the β-tocopherol content is preferably 5 or more, and the mass ratio (γ / δ) of the γ-tocopherol content to the δ-tocopherol content is preferably 8.8 or less.SELECTED DRAWING: Figure 2

Description

Technical Field

[0001] The present invention relates to a nutritional composition containing oligosaccharides and tocopherols.

Background Art

[0002] It is known that Bifidobacterium bacteria are the most dominant in the intestinal flora of infants. Bifidobacterium bacteria are obligate anaerobic bacteria that inhabit the lower small intestine to the large intestine of healthy humans, and are widely known to play an important role in maintaining the health of the host through functions such as intestinal regulation and immune regulation (Non-Patent Document 1). Therefore, developing foods that promote the growth of Bifidobacterium bacteria is important for maintaining the health of infants.

[0003] In recent years, it has been considered that a sugar source is particularly important for the growth of Bifidobacterium bacteria, and various oligosaccharides themselves and sugar sources combining various oligosaccharides with other components have been proposed for use as so-called "prebiotics".

[0004] Among oligosaccharides, human milk oligosaccharides (hereinafter also referred to as HMO), which are a general term for various oligosaccharides contained in human colostrum, have also been studied variously. Human milk oligosaccharides are calculated to be contained at 22 - 24 g / L in colostrum and 12 - 13 g / L in mature milk, and are the third solid component after lactose and lipids in breast milk. Human milk oligosaccharides are known to have functions such as infection defense and immune regulation, and it has also been reported that they are selectively utilized by Bifidobacterium bacteria in the intestine of infants and promote their growth (Patent Document 1).

[0005] In addition, tocopherol has attracted attention as a component to be blended in nutritional compositions for infants, and it has been reported that α-tocopherol promotes the growth of Bifidobacterium bacteria (Patent Document 2). In addition, Patent Document 3 discloses a composition containing 2'-fucosyllactose, which is one of human milk oligosaccharides, and α-tocopherol, and it is described that such a composition has an effect of improving at least one of cognition, learning, and memory. There are no findings as described above regarding the isomers of tocopherol other than α-tocopherol.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0007]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0008] An object of the present invention is to provide a technique for further promoting the growth of Bifidobacterium bacteria in the intestinal flora.

Means for Solving the Problems

[0009] As a result of intensive studies to solve the above problems, the present inventors have found that by combining oligosaccharides such as human milk oligosaccharides with isomers of tocopherol other than α-tocopherol, the growth of Bifidobacterium bacteria can be further promoted, and thus the present invention has been completed.

[0010] That is, the present invention is a nutritional composition containing oligosaccharide and tocopherol, wherein the tocopherol contains one or more selected from the group consisting of β-tocopherol, γ-tocopherol, and δ-tocopherol (excluding breast milk). In the nutritional composition of the present invention, the mass ratio (δ / β) of the content of δ-tocopherol to the content of β-tocopherol is preferably 5 or more, more preferably 10 or more. In the nutritional composition of the present invention, the mass ratio (γ / δ) of the content of γ-tocopherol to the content of δ-tocopherol is preferably 8.8 or less, more preferably 3.9 or less. In the nutritional composition of the present invention, the mass ratio of the total content of β-tocopherol, γ-tocopherol, and δ-tocopherol to the content of oligosaccharide is preferably 0.001 to 0.2. In the nutritional composition of the present invention, the total content of β-tocopherol, γ-tocopherol, and δ-tocopherol with respect to the whole composition is preferably 0.0002 to 0.1% by mass. In the nutritional composition of the present invention, the content of oligosaccharide with respect to the whole composition is preferably 0.05 to 10% by mass. In the nutritional composition of the present invention, the oligosaccharide preferably has 3 to 5 sugar residues. Further, the oligosaccharide is preferably human milk oligosaccharide, more preferably one or more selected from the group consisting of lacto-N-neotetraose (also denoted as LNnT), lacto-N-tetraose (also denoted as LNT), 2'-fucosyllactose (also denoted as 2'-FL), 3'-sialyllactose (also denoted as 3'-SL), 6'-sialyllactose (also denoted as 6'-SL), difucosyllactose (also denoted as DFL), and 3-fucosyllactose (also denoted as 3-FL). The nutritional composition of the present invention is suitable for use in promoting the growth of Bifidobacterium bacteria, and the Bifidobacterium bacteria are preferably one or more selected from the group consisting of Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium longum. The nutritional composition of the present invention preferably further contains Bifidobacterium bacteria. The nutritional composition of the present invention is preferably a prepared milk.

Effects of the Invention

[0011] According to the present invention, there is provided a nutritional composition capable of promoting the growth of Bifidobacterium bacteria in the intestine. Such a nutritional composition can be in the form of an oral composition such as a food or drink or a pharmaceutical, or can be contained in a food or drink or a pharmaceutical in the form of an additive or the like. By ingesting the nutritional composition of the present invention, an effect of improving the intestinal flora can be obtained, and it is expected to be useful for maintaining the health of the consumer, particularly infants.

Brief Description of the Drawings

[0012]

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Mode for Carrying Out the Invention

[0013] Next, the present invention will be described in detail. However, the present invention is not limited to the following embodiments and can be freely changed within the scope of the present invention.

[0014] The nutritional composition of the present invention contains oligosaccharide and tocotrienol. However, breast milk itself is not included in the nutritional composition of the present invention. The tocotrienol includes one or more selected from the group consisting of β-tocotrienol, γ-tocotrienol, and δ-tocotrienol. Preferably, it includes all three of β-tocotrienol, γ-tocotrienol, and δ-tocotrienol. Further, the nutritional composition of the present invention may further contain α-tocotrienol.

[0015] In the nutritional composition of the present invention, the mass ratio (δ / β) of the content of δ-tocotrienol to the content of β-tocotrienol is preferably 5 or more, more preferably 10 or more. The upper limit of δ / β is not particularly limited, but is usually 200. By setting δ / β within the above range, the growth promoting effect on Bifidobacterium bacteria can be more easily obtained.

[0016] In the nutritional composition of the present invention, the mass ratio (γ / δ) of the content of γ-tocotrienol to the content of δ-tocotrienol is preferably 8.8 or less, more preferably 3.9 or less. The lower limit of γ / δ is not particularly limited, but is usually 0.1. By setting γ / δ within the above range, the growth promoting effect on Bifidobacterium bacteria can be more easily obtained.

[0017] In the nutritional composition of the present invention, the mass ratio of the total content of β-tocopherol, γ-tocopherol and δ-tocopherol to the content of oligosaccharides is preferably 0.001 to 0.2, more preferably 0.004 to 0.2, and still more preferably 0.01 to 0.2. By setting the mass ratio within the above range, the growth promoting effect on Bifidobacterium bacteria can be more easily obtained.

[0018] In the nutritional composition of the present invention, the total content of β-tocopherol, γ-tocopherol and δ-tocopherol with respect to the whole composition is preferably 0.0002 to 0.1% by mass, more preferably 0.001 to 0.04% by mass, and still more preferably 0.002 to 0.04% by mass. By setting the total content within the above range, the growth promoting effect on Bifidobacterium bacteria can be more easily obtained.

[0019] In the nutritional composition of the present invention, the total content of oligosaccharides is preferably 0.05 to 10% by mass, more preferably 0.1 to 10% by mass, and still more preferably 0.2 to 10% by mass with respect to the whole composition. By setting the total content within the above range, the growth promoting effect on Bifidobacterium bacteria can be more easily obtained.

[0020] The oligosaccharides in the present invention refer to oligosaccharides in which sugars of two or more sugars are bonded by glycosidic bonds. As the oligosaccharides in the present invention, the number of sugar residues bonded to each other by glycosidic bonds is 2 or more, preferably 2 or more and 10 or less, and particularly preferably 3 or more and 5 or less. Examples of the oligosaccharides in the present invention include, but are not limited to, raffinose, lactulose, maltotriose, stachyose, galactooligosaccharide, fructooligosaccharide, kestose, nystose, soy oligosaccharide, lactulose oligosaccharide, xylooligosaccharide, isomaltooligosaccharide, coffee bean manno-oligosaccharide, gluconic acid, polydextrose, inulin, etc. Further, it may contain one or more of the above-described oligosaccharides. Among these, as oligosaccharides having 3 to 5 sugar residues, maltotriose, stachyose, galactooligosaccharide, fructooligosaccharide, kestose, nystose, and human milk oligosaccharide are preferable.

[0021] Human milk oligosaccharide is particularly preferably mentioned as the oligosaccharide in the present invention. The human milk oligosaccharide is not particularly limited as long as it is an oligosaccharide usually contained in human milk, and examples thereof include fucosylated oligosaccharide, sialylated oligosaccharide, acetylated oligosaccharide, galactosyllactose, etc. Examples of the fucosylated oligosaccharide include 2'-fucosyllactose, lacto-N-fucopentaose-I, lacto-N-fucopentaose-II, lacto-N-fucopentaose-III, etc. Examples of the sialylated oligosaccharide include 3'-sialyllactose, 6'-sialyllactose, difucosyllactose, 3-fucosyllactose, lactodifucotetraose, difucosyllacto-N-tetraose, sialyllacto-N-tetraose a, sialyllacto-N-tetraose b, sialyllacto-N-tetraose c, disialyllacto-N- tetraose, etc. Examples of the acetylated oligosaccharide include lacto-N-neotetraose, lacto-N-tetraose, lacto-N-hexaose, etc. Examples of the galactosyllactose include 3'-galactosyllactose, 4'-galactosyllactose, 6'-galactosyllactose, etc. Among these, from the viewpoint of the growth promoting effect on Bifidobacterium bacteria, one or more selected from lacto-N-neotetraose, lacto-N-tetraose, 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, difucosyllactose, and 3-fucosyllactose are particularly preferred.

[0022] In addition, the human milk oligosaccharide in the present invention may be purified or a mixture as long as the effects of the present invention are not impaired. Further, it may contain one or more of the above-described human milk oligosaccharides.

[0023] The nutritional composition of the present invention can promote the growth of bacteria belonging to the genus Bifidobacterium. More specifically, while oligosaccharides are usually assimilated by Bifidobacterium bacteria and promote their growth, β-tocopherol, γ-tocopherol, and / or δ-tocopherol can enhance the growth promoting effect, thereby becoming excellent prebiotics. The Bifidobacterium bacteria whose growth is promoted by the nutritional composition of the present invention are not particularly limited. For example, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis (reclassified as Bifidobacterium longum subsp. infantis), Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium animalis , Bifidobacterium pseudolongum, Bifidobacterium reuteri, Bifidobacterium catenulatum , Bifidobacterium pseudocatenulatum, Bifidobacterium lactis etc. Among these, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum are four types that are particularly important for the health of the intestinal flora in infancy (so-called "infant-type bifidobacteria"), and it is preferable that their growth is promoted.

[0024] In this specification, "growth" of bacteria includes an increase in the absolute number of bacteria, and "promotion of growth" means that the degree of such increase is greater when the nutritional composition of the present invention is applied than when it is not applied. That is, when the nutritional composition of the present invention is applied in vivo or in vitro, the number of bacteria of the genus Bifidobacterium increases compared to when it is not applied. The degree of increase in the number of such bacteria is not particularly limited, but is preferably 1.1 times or more, more preferably 1.5 times or more, and even more preferably 3 times or more the number of bacteria compared to the number of such bacteria when the nutritional 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, for example, by measuring the turbidity (absorbance) of the digestive tract contents of animals such as humans who have ingested the medium or composition in which the bacteria are cultured or the amount of short-chain fatty acids such as acetic acid and observing an increase in the value, or by measuring the pH in the medium and observing a decrease in the value.

[0025] In addition, "proliferation" includes an increase in the proportion of the bacteria in the intestinal flora, and "promotion of proliferation" means that the degree of such increase is greater than when the nutritional composition of the present invention is not applied. That is, it includes increasing the proportion of Bifidobacterium bacteria present in the digestive tract of animals such as humans who have ingested the nutritional composition of the present invention. Here, the "proportion" can also be rephrased as the "occupancy rate" with respect to the entire bacterial flora detected in the intestinal flora. The "increase in proportion" may simultaneously occur with an increase or decrease in the proportion of other bacteria in the intestinal flora as long as the proportion of Bifidobacterium bacteria in the intestinal flora increases. The degree of such increase in proportion is not particularly limited, but it preferably means that it is 2% or more, more preferably 5% or more, and still more preferably 20% or more greater than the proportion of the bacteria when the nutritional composition of the present invention is not applied.

[0026] In addition, "promotion of proliferation" may be 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 such increase in rate is not particularly limited, but it preferably means that it is 10% or more, more preferably 20% or more, and still more preferably 50% or more greater than the rate of the bacteria when the nutritional composition of the present invention is not applied. The increase in rate can be confirmed, for example, by the fact that the time required to reach an arbitrary number of bacteria or proportion is shorter when the nutritional composition of the present invention is applied than when it is not applied.

[0027] The nutritional composition of the present invention may itself be in the form of food and drink, pharmaceuticals, etc., or may be in the form of being contained in food and drink, pharmaceuticals, etc. as an additive. The ingestion (administration) route of the nutritional composition of the present invention may be either oral or parenteral, but is usually oral. Examples of parenteral ingestion (administration) include rectal administration.

[0028] As described above, since the nutritional composition of the present invention can promote the growth of Bifidobacterium bacteria, it is preferable to include Bifidobacterium bacteria in the composition together with one or more tocopherols selected from the group consisting of oligosaccharides and β-tocopherol, γ-tocopherol, and δ-tocopherol. Here, the Bifidobacterium bacteria to be included in the nutritional composition include Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium pseudolongum, Bifidobacterium reuteri, Bifidobacterium catenulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium lactis, etc., and are not particularly limited. However, it is preferable to include Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum, which are particularly important in the above-mentioned infants.

[0029] As Bifidobacterium longum, Bifidobacterium longum NITE BP-02621 (alias: BB536 or Bifidobacterium longu m subsp. longum ATCC BAA-999) can be used. Bif idobacterium longum BB536 was deposited internationally under the Budapest Treaty with the accession number of NITE BP-02621 at the NPMD on January 26, 2018. There is. Bifidobacterium longum subsp. longum ATCC BAA-999 (number: ATCC BAA-999), which is the same bacterium as this, is A merican Type Culture Collection (ATCC: United States, 2 0110, Manassas, Virginia, University Boulevard 10801 (108 01 University Boulevard, Manassas, VA 2011 It is available as ATCC BAA-999 from ((0, United States of America)) (see, for example, JP-A-2012-223134, etc.). In addition, as Bifidobacterium longum, Bifidobacterium longum available from ATCC can be used. Bifidobacterium longum subsp. longum ATCC 15707 can be used. It can be used.

[0030] Examples of Bifidobacterium breve include Bifidobacterium breve M-1 6V. Bifidobacterium breve M-16V was deposited on January 26, 2018, with the Patent Microorganisms Depositary, National Institute of Technology and Evaluation (NPMD) (Room 122, 2-5-8 Kazusa Kamashima, Kisarazu City, Chiba Prefecture 292-0818, Japan) under the Budapest Treaty with the accession number NITE BP-02622. It may be used as a commercially available product, for example, "Bifidobacterium breve M-16 V" manufactured by Morinaga Milk Industry Co., Ltd. It may also be used. V". Examples of Bifidobacterium breve also include Bifidobacterium breve MCC1274. Bifidobacterium breve MCC1274 was deposited on August 25, 2009, with the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology (currently the Patent Organism Depositary, National Institute of Technology and Evaluation (IPOD) (Room 120, 2-5-8 Kazusa Kamashima, Kisarazu City, Chiba Prefecture 292-0818, Japan) under the Budapest Treaty with the accession number FERM BP-1 1175. An international deposit based on the Budapest Treaty has been made.

[0031] Examples of Bifidobacterium infantis include Bifidobacterium longum subsp. infantis MCC2042. Bifidobacterium longum subsp. infantis MCC2042 was deposited with the NPMD on November 20, 2019, under the accession number NITE BP-03068, in accordance with the Budapest Treaty for international deposit. Examples of Bifidobacterium infantis also include Bifidobacterium infantis M-63. Bifidobacterium infantis M-63 was deposited with the NPMD on January 26, 2018, under the accession number NITE BP-02623, in accordance with the Budapest Treaty for international deposit. In addition, as Bifidobacterium infantis, Bifidobacterium longum subsp. infantis ATCC15697, which is available from ATCC, can be used.

[0032] Examples of Bifidobacterium bifidum include Bifidobacterium bifidum MCC2030. Bifidobacterium bifidum MCC2030 was deposited with the NPMD on November 8, 2019, under the accession number NITE BP-03058, in accordance with the Budapest Con vention for international deposit. Examples of Bifidobacterium bifidum also include Bifidobacterium bifidum MCC1092. Bifidobacterium bifidum MCC1092 was deposited with the NPMD on February 21, 2017, under the accession number NITE BP-02429, in accordance with the Budapest Con vention for international deposit. In addition, as Bifidobacterium bifidum, Bifidobacterium bifidum MCC1319 can be mentioned. Bifidobacterium bifidum MCC1319 was deposited with the NPMD on February 21, 2017, under the accession number NITE BP-02431, in accordance with the Budape st Treaty for international deposit. In addition, examples of Bifidobacterium bifidum include Bifidobacterium bifidum MCC1868. Bifidobacterium bifidum MCC1868 was internationally deposited with NPMD on February 21, 2017, under the accession number NITE BP-02432, in accordance with the Budapest Treaty. In addition, examples of Bifidobacterium bifidum include Bifidobacterium bifidum MCC1870. Bifidobacterium bifidum MCC1870 was internationally deposited with NPMD on February 21, 2017, under the accession number NITE BP-02433, in accordance with the Budapest Treaty.

[0033] Note that the bacteria identified by the above-exemplified bacterial names are not limited to the strains themselves that have been deposited or registered with a predetermined institution under the said bacterial names (hereinafter, also referred to as "deposited strains" for convenience of explanation), but also include strains that are substantially equivalent thereto ("derived strains" or "induced strains"). That is, it is not limited to the strains themselves deposited with the above depository institution under the above accession numbers, but also includes strains that are substantially equivalent thereto. For each bacterium, "a strain that is substantially equivalent to the above deposited strain" means a strain that belongs to the same species as the above deposited strain, and the nucleotide sequence of its 16S rRNA gene has, preferably, 99.86% or more, more preferably 99.93% or more, and even more preferably 100% identity to the nucleotide sequence of the 16S rRNA gene of the above deposited strain, and preferably has the same mycological properties as the above deposited strain. For each bacterium, a strain that is substantially equivalent to the above deposited strain may be, for example, a derived strain having the above deposited strain as the parent strain. Examples of derived strains include strains bred from the deposited strain and strains naturally occurring from the deposited strain. Breeding methods include modification by genetic engineering techniques and modification by mutagenesis treatment. Mutagenesis treatment includes irradiation with X-rays, irradiation with ultraviolet rays, and N-methyl-N'-nitro-N-nitrosoguanidine Examples of treatment with mutagens such as ethyl methanesulfonate and methyl methanesulfonate can be mentioned. Examples of strains that occur naturally from the deposited strain include strains that occur naturally when using the deposited strain. Such strains include mutant strains that occur naturally by culturing the deposited strain (for example, subculture). The derived strain may be constructed by one kind of modification, or may be constructed by two or more kinds of modifications.

[0034] As the cells of the genus Bifidobacterium to be contained in the nutritional composition of the present invention, commercially available products may be used, or those appropriately produced and obtained may be used. In addition, the cells of the genus Bifidobacterium to be contained in the nutritional composition of the present invention can be easily obtained by culturing the aforementioned genus Bifidobacterium. The culturing method is not particularly limited as long as the genus Bifidobacterium can grow. As the culturing method, for example, the method usually used for culturing the genus Bifidobacterium can be used as it is or after being appropriately modified. The culturing temperature may be, for example, 25 to 50°C, and preferably 35 to 42°C. The culturing can preferably be carried out under anaerobic conditions, for example, while aerating an anaerobic gas such as carbon dioxide gas. In addition, the culturing can also be carried out under microaerobic conditions such as liquid static culture. The culturing can be carried out, for example, until the genus Bifidobacterium grows to a desired degree or for 5 to 24 hours.

[0035] The medium used for cultivation is not particularly limited as long as Bifidobacterium bacteria can grow. As the medium, for example, a medium commonly 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, and molasses can be used according to their assimilability. As the nitrogen source, for example, ammonium salts and nitrates such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate can be used. In addition, as inorganic salts, for example, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, ferrous sulfate, etc. can be used. Also, organic components such as peptone, soybean powder, defatted soybean meal, meat extract, and yeast extract, and antioxidants such as ascorbic acid or its derivatives or their salts, and α-lipoic acid or its derivatives or their salts may be used. Specific examples of the medium commonly used for culturing Bifidobacterium bacteria include Reinforced Clostridial medium, MRS medium (de Man, Rogosa, and Sharpe medium), mMRS medium (modified MRS medium), TOSP medium (TOS propionate medium), and TOSP Mup medium (TOS propionate mupirocin medium).

[0036] As the Bifidobacterium bacteria that can be contained in the nutritional composition of the present invention, the bacterial cells or the fraction containing the same can be used without particular limitation. That is, as the Bifidobacterium bacteria, for example, the culture obtained by culturing may be used as it is, the culture may be diluted or concentrated and used, or the bacterial cells recovered from the culture may be used. Further, as long as the effect of improving the intestinal flora is not impaired, various additional operations such as heating and freeze-drying can be performed after culturing. The additional operation is preferably one with high viability of the bacterial cells. That is, as the Bifidobacterium bacteria that can be contained in the nutritional composition of the present invention, specifically, the culture of Bifidobacterium bacteria, or the bacterial cells recovered from the culture, and the processed products thereof can be mentioned. Examples of the processed products include dilutions, concentrates, or dried products. Note that the bacterial cells are preferably used in a form containing viable bacterial cells. The bacterial cells may be composed of viable bacterial cells, for example, or may be a mixture of viable bacterial cells and dead bacterial cells. To give an example of the acquisition procedure of the Bifidobacterium bacteria that can be contained in the nutritional composition of the present invention, Bifidobacterium bacteria are inoculated into a medium containing a carbon source, a nitrogen source, inorganic salts, an organic component, and ascorbic acid in an amount of 0.1 to 2.0% by mass based on the whole medium, cultured at 35 to 42 ° C for 5 to 24 hours, the bacterial cells are collected from the culture by centrifugation, and the recovered wet bacterial cells are freeze-dried to obtain a bacterial powder. According to such a procedure, it is preferable because a bacterial powder of Bifidobacterium bacteria with high storage stability of viable bacterial cells can be obtained.

[0037] In the nutritional composition of the present invention, the Bifidobacterium bacteria contained in the composition contain at least viable bacteria, and in the composition, in total, 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, even more preferably 1×10 6 ~1×10 11cfu / g or 1×10 6 ~1×10 11 cfu / mL contains viable bacteria of the genus Bifidobacterium. In addition, in the nutritional composition of the present invention, the total viable count of the genus Bifidobacterium bacteria contained in the composition is preferably 1×10 per 1 g of oligosaccharide contained in the composition 8 ~1×10 11 cfu, more preferably 1×10 8 ~1×10 10 cfu, even more preferably 1×10 8 ~3×10 9 cfu. As long as viable bacteria are contained, dead bacteria may also be included. Here, cfu refers to Colony forming unit. In this specification, for example, it can be taken as the value when cultured at 38 °C on a solid medium containing 10% by mass of reduced skim milk powder. These ranges may be the content when distributed as a nutritional composition or the content when orally ingested.

[0038] As described above, since the nutritional composition of the present invention can promote the growth of bacteria of the genus Bifidobacterium such as Bifidobacterium infantis, it can be preferably applied to the use of promoting the growth of bacteria of the genus Bifidobacterium.

[0039] Here, the bacteria of the genus Bifidobacterium to be grown are not limited to the bacteria of the genus Bifidobacterium contained in the nutritional composition of the present invention, but also include bacteria of the genus Bifidobacterium present in the digestive tract of animals such as humans who have ingested orally. Therefore, the nutritional composition of the present invention can be used to improve the intestinal flora. Here, "flora improvement" includes increasing the number of bacteria and their proportion present in the intestinal flora of Bifidobacterium bacteria, particularly Bifidobacterium infantis. Further, "flora improvement" may include increasing the proportion of other beneficial bacteria present in the intestinal flora as long as the proportion of Bifidobacterium bacteria in the intestinal flora increases, or decreasing the proportion of harmful bacteria present in the intestinal flora. Examples of other beneficial bacteria include lactic acid bacteria. Examples of harmful bacteria include Clostridium welchii, Salmonella bacteria, Staphylococcus aureus, pathogenic Escherichia coli, and the like. The "proportion" can also be referred to as the "occupancy rate" with respect to the entire bacterial flora detected in the intestinal flora.

[0040] The nutritional 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 subjects with diseases or conditions caused by deterioration of the intestinal flora. For example, it can be used for intestinal regulation, immune regulation, anti-allergy, prevention of bacterial and viral infections, reduction of oxidative stress, prevention and improvement of diarrhea, prevention and improvement of constipation, inflammatory bowel disease, prevention of colorectal cancer, and the like.

[0041] The present invention can be rephrased as the use of oligosaccharides and one or more tocopherols selected from the group consisting of β-tocopherol, γ-tocopherol, and δ-tocopherol in the production of a composition for promoting the growth of Bifidobacterium bacteria. The present invention can also be rephrased as the use of oligosaccharides and one or more tocopherols selected from the group consisting of β-tocopherol, γ-tocopherol, and δ-tocopherol in promoting the growth of Bifidobacterium bacteria. The present invention can also be rephrased as oligosaccharides and one or more tocopherols selected from the group consisting of β-tocopherol, γ-tocopherol, and δ-tocopherol used to promote the growth of Bifidobacterium bacteria.

[0042] The present invention can also be described as a method for promoting the growth of Bifidobacterium bacteria, which includes administering to a subject one or more tocopherols selected from the group consisting of oligosaccharides and β-tocopherol, γ-tocopherol, and δ-tocopherol. 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 "causing a subject to ingest". The ingestion may be voluntary (free ingestion) or forced (forced ingestion). That is, specifically, the administration step may be, for example, a step of formulating one or more tocopherols selected from the group consisting of oligosaccharides and β-tocopherol, γ-tocopherol, and δ-tocopherol into food and drink products or feed and supplying them to the subject, thereby allowing the subject to freely ingest the food and drink products or feed.

[0043] The timing of ingestion (administration) of the nutritional composition of the present invention is not particularly limited and can be appropriately selected according to the state of the administration subject.

[0044] The amount of ingestion (administration) of the nutritional composition of the present invention is appropriately selected according to the age, sex, state, and other conditions of the ingestion (administration) subject. Note that regardless of the amount and duration of ingestion (administration), the drug can be administered once a day or divided into multiple times.

[0045] The nutritional composition of the present invention is preferably in the form of food and drink products. As food and drink products, as long as they do not impair the effects of the present invention, the form and properties are not particularly limited, and they can be produced by ordinary methods using raw materials commonly used in food and drink products. Note that the form of additives added to food and drink products or pharmaceuticals is also included in the nutritional composition of the present invention. Examples of such forms include additives added to expressed breast milk or formula milk, and it is assumed that the milk after addition is ingested by newborns and infants. Food and drink products are usually ingested orally, but are not limited to this. For example, they may be ingested nasally, or through a gastrostomy or enterostomy. For example, it is envisaged that for newborns and infants, prepared milk, which is the nutritional composition of the present invention described below, or breast milk added with the nutritional composition of the present invention, will be ingested through a nasogastric feeding tube or the like.

[0046] Food and drink products include, regardless of their form such as liquid, paste, gel-like solid, powder, etc. For example, confectionery; wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, tempura flour, breadcrumbs, etc.; instant foods such as instant noodles, cup noodles, retort foods, canned cooked foods, microwave foods, instant soups and stews, instant miso soup and drinks, canned soups, freeze-dried foods, and other instant foods; agricultural processed products such as canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, cereals (processed grain products), etc.; fishery processed products such as canned fishery products, fish ham and sausage, fish paste products, seafood delicacies, tsukudani, etc.; livestock processed products such as canned livestock products and pastes, livestock meat ham and sausage, etc.; milk and dairy products such as processed milk, milk beverages, yogurts, lactic acid bacteria beverages, cheese, ice creams, cream, and other dairy products; fats and oils such as butter, margarines, vegetable oils, etc.; basic seasonings such as soy sauce, miso, sauces, processed tomato seasonings, mirin, vinegars, etc.; compound seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other compound seasonings; frozen foods such as raw material frozen foods, semi-cooked frozen foods, cooked frozen foods, etc.; confectionery such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese confectionery, rice confectionery, bean confectionery, dessert confectionery, jelly, and other confectionery; soft drinks, natural fruit juices, fruit juice beverages, soft drinks with fruit juice, fruit pulp beverages, fruit drinks with fruit pieces, vegetable-based beverages, soy milk, soy milk beverages, coffee beverages, tea beverages, powdered beverages, concentrated beverages, sports beverages, nutritional beverages, alcoholic beverages, and other preferred beverages; baby food, furikake, nori for ochazuke, and other commercially available foods; nutritional compositions such as prepared milk (including powdered milk, liquid milk, etc.), liquid foods, supplements, etc.; functional foods (foods for specified health use, foods with nutritional functions), etc.

[0047] Among these, nutritional compositions are preferably mentioned. In the present invention, the "nutritional composition" is not particularly limited as one aspect of food and drink, but is preferably prepared milk, liquid food, supplements, etc., and more preferably prepared milk. The ingestion target is not limited to infants, young children, children, and adults, but is preferably infants and young children. Prepared milk includes prepared powdered milk and prepared liquid milk. Prepared powdered milk is defined in the ordinance of the Ministry of Health, Labour and Welfare regarding the ingredient standards of milk and dairy products (Ordinance on Milk etc.) as "raw milk, cow's milk, special cow's milk, or foods manufactured using these as raw materials, processed, or made into a powder form with the addition of nutrients necessary for infants as the main raw material". Prepared liquid milk is defined in the above ordinance as "raw milk, cow's milk, special cow's milk, or foods manufactured using these as raw materials, processed, or made into a liquid form with the addition of nutrients necessary for infants as the main raw material". In addition, prepared milk is a product in which various nutritional components such as various proteins, fats and oils, carbohydrates, minerals, and vitamins are blended, and also includes those processed into a powder form or a liquid form. In addition, prepared milk further includes "prepared powdered milk for infants", "prepared liquid milk for infants", and "powdered milk for pregnant and lactating women" in the foods for special dietary uses defined by the Health Promotion Act, and also includes aspects such as prepared powdered milk for young children, nutritional powder for adults, and nutritional powder for the elderly.

[0048] When the nutritional 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.; and so on. In such formulation, it can follow the description of the components, carriers, and methods related to the formulation of pharmaceuticals described later.

[0049] In addition, it can also be used as feed as one aspect of food and drink. Examples of feed include pet food, livestock feed, fish feed, etc. The form of the feed is not particularly limited. For example, grains such as corn, wheat, barley, rye, and milo; vegetable oil meals such as soybean meal, rapeseed meal, coconut meal, and linseed meal; bran such as bran, wheat bran, rice bran, and defatted rice bran; manufacturing meals such as corn gluten meal and corn germ meal; animal feeds such as fish meal, skim milk powder, whey, yellow grease, and tallow; Torula yeast Yeasts such as yeast and brewer's yeast; mineral feeds such as tricalcium phosphate and calcium carbonate; oils and fats; single amino acids; sugars, etc. may be contained.

[0050] When the nutritional 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 with the indication of the use of growing Bifidobacterium bacteria in the intestine.

[0051] Such "indication" acts include all acts for informing consumers of the above-mentioned use. As long as it is an expression that can make consumers recall or infer the above-mentioned use, regardless of the purpose of the indication, the content of the indication, the object / media to be indicated, etc., all fall within the "indication" acts of the present invention. In addition, the "indication" is preferably made by an expression that allows consumers to directly recognize the above-mentioned use. Specifically, acts such as transferring, delivering, displaying for transfer or delivery, importing a product related to a food or drink or its packaging with the above-mentioned use described thereon, advertising related to the product, displaying, distributing, or publishing a price list or transaction document with the above-mentioned use described thereon, or providing information containing these by electromagnetic (such as the Internet) methods with the above-mentioned use described thereon.

[0052] On the other hand, as the indication content, it is preferably an indication approved by the administration or the like (for example, an indication made in a manner based on various systems determined by the administration and approved based on such approval). It is also preferable to attach such indication content to promotional materials at the point of sale such as packaging, containers, catalogs, pamphlets, POPs, and other documents.

[0053] In addition, "labeling" includes labeling as health foods, functional foods, enteral nutrition foods, special-purpose foods, health-functional foods, foods for specified health use, nutritional-functional foods, functional-labeling foods, quasi-drugs, etc. Among these, in particular, labeling approved by the Consumer Affairs Agency, for example, labeling approved under the systems related to foods for specified health use, nutritional-functional foods, or functional-labeling foods, or labeling approved under similar systems, etc. are included. Specifically, labeling as a food for specified health use, labeling as a conditional food for specified health use, labeling indicating an impact on the structure and function of the body, disease risk reduction labeling, functional labeling based on scientific evidence, etc. can be mentioned. More specifically, labeling as a food for specified health use (particularly labeling of the use for health) and similar labeling defined in the Cabinet Office Ordinance (Cabinet Office Ordinance No. 57 of August 31, 2009) regarding permission for special-purpose labeling stipulated in the Health Promotion Act are typical examples. Examples of such labeling include labeling such as "For those who want to increase Bifidobacterium", "To increase Bifidobacterium useful for the health of babies", "To improve the intestinal flora", "To improve the gut microbiota", "For the health of the stomach of infants", etc.

[0054] When the nutritional composition of the present invention is in the form of a pharmaceutical, the administration route may be either oral or parenteral, but oral administration is preferred. In addition, examples of parenteral intake (administration) include rectal administration, etc. As the form of the pharmaceutical, it can be formulated into a desired dosage form as appropriate according to the administration method. For example, in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets, capsules, etc.; liquid preparations such as solutions, syrups, suspensions, emulsions, etc. In addition, in the case of parenteral administration, it can be formulated into suppositories, ointments, injections, etc. When formulating, components such as excipients, pH adjusters, colorants, flavoring agents, etc. that are usually used in formulation can be used. In addition, it is also possible to use in combination other medicinal ingredients, prebiotics for Bifidobacterium genus bacteria that are known or will be found in the future, prebiotics for other bacteria, etc. In addition, the formulation can be carried out by a method known per se according to the dosage form. When formulating, a formulation carrier may be appropriately blended for formulation.

[0055] 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, hydroxypropyl methylcellulose, carboxymethyl cellulose, and carboxymethyl cellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium metasilicate aluminate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; sulfate derivatives such as calcium sulfate and the like.

[0056] Examples of binders include, for example, in addition to the above excipients, gelatin; polyvinylpyrrolidone; macrogol and the like.

[0057] Examples of disintegrants include, for example, in addition to the above excipients, chemically modified starches or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone.

[0058] Examples of lubricants include, for example, talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as beeswax and carnauba wax; 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; silicas such as anhydrous silicic acid and hydrated silicic acid; starch derivatives and the like.

[0059] Examples of stabilizers include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid, and the like.

[0060] Examples of flavoring and odor-masking agents include sweeteners, acidulants, fragrances, and the like. In addition, examples of carriers used in the case of liquid preparations for oral administration include solvents such as water.

[0061] The timing of ingesting the pharmaceutical of the present invention is not particularly limited, for example, before meals, after meals, between meals, before bedtime, and the like.

Examples

[0062] The present invention will be described in more detail below using examples, but the present invention is not limited to these examples.

[0063] <Test Example 1> Evaluation of the effect of promoting the growth of Bifidobacterium bacteria by single culture of each strain (1) Single culture Bifidobacterium infantis M-63, Bifidobacterium infantis MCC2042, or Bifidobacterium bifidum MCC2030 was cultured in MRS medium at 37°C for 16 hours to prepare a preculture solution. ​The components shown in Table 1 (except for cysteine, tocopherol, and oligosaccharides) were dissolved in purified water to prepare a medium with double the concentration, and the pH was adjusted to 6.5, followed by autoclaving at 121°C for 15 minutes. Filter-sterilized cysteine ​​was then added. Tocopherol was dissolved in dimethyl sulfoxide (Fujifilm Wako Pure Chemical Industries, Ltd.) and oligosaccharides in purified water, respectively, and filter-sterilized and added to the above medium. The concentrations were then adjusted with sterilized purified water to the final concentrations shown in Table 1. 200 μL of the prepared medium was added to a 96-well plate (n=6 per group), and then Bifidobacterium infantis M-63, Bifidobacterium infantis M-64, Bifidobacterium infantis M-65, Bifidobacterium infantis M-66, Bifidobacterium infantis M-67, Bifidobacterium infantis M-68, Bifidobacterium infantis M-69 ... The preculture solution of Bifidobacterium bifidum MCC2030 or Fantis MCC2042 was added to the medium at 1% of the medium volume. The medium was cultured in an anaerobic chamber (Baker Ruskinn) at 37°C for 16 hours, and the turbidity (OD 600 ) was measured.

[0064] [Table 1-1]

[0065] [Table 1-2]

[0066] [Table 1-3]

[0067] [Table 1-4]

[0068] (2) Results The difference in turbidity from the start of cultivation to 16 hours after cultivation (ΔOD 600) is shown in FIGS. 1 to 12 as the growth degree of each strain of Bifidobacterium bacteria. In comparison with the case where any strain contains no tocopherol but contains human milk oligosaccharide, the growth of Bifidobacterium bacteria was more promoted when it contained a combination of three kinds of tocopherols and human milk oligosaccharide. In addition, when it contained three kinds of tocopherols but no human milk oligosaccharide, no growth of Bifidobacterium bacteria was observed.

[0069] <Test Example 2> Evaluation of the effect of promoting the growth of Bifidobacterium bacteria by fecal culture (1) Pretreatment of samples Fecal samples were obtained from two healthy infants aged 1.5 or 7 months. They were collected immediately after defecation and immediately transferred to a temperature of 10°C or lower under anaerobic conditions. Within 8 hours after defecation, the collected feces were diluted with 10 times the volume of physiological saline and stored at -80°C until use.

[0070] (2) Fecal culture In order to simulate the environment in the large intestine, a culture experiment was conducted according to the following procedure. A Bio Jr.8 100 mL × 8 multi-culture device (BJR-25NA1S-8M manufactured by Biott Co., Ltd.) was used, and the operation was in accordance with the equipment manual. As the culture medium, 100 mL of YCFA medium with the composition shown in Table 2 was used. The medium components other than tocopherol and oligosaccharide were dissolved in purified water and sterilized by autoclaving. Tocopherol was dissolved in dimethyl sulfoxide (DMSO, manufactured by Fujifilm Wako Pure Chemical Corporation), and oligosaccharide was dissolved in purified water and then filter-sterilized, and then added to the autoclaved product. The culture temperature was 37°C, filtered CO2 was blown into each culture container to maintain an anaerobic state during the culture period. After adjusting the pH to 7.0, 100 μL (10 mg as feces) of the fecal sample diluted with physiological saline was added to start anaerobic culture. In order to simulate the pH in the large intestine of infants, when the pH dropped below 5.5 during the culture period, it was neutralized with 1M Na2CO3, and the Control was carried out. The medium was collected 7 hours after the start of culture, centrifuged at 8000 rmp, 3 min, and 4°C, and the supernatant and precipitate were collected respectively.

[0071] (3) DNA Extraction and Bacterial Count Measurement DNA was extracted from the precipitate recovered in (2 ) using the GENE PREP STAR PI-480 Automatic Extractor (Kurabo Industries Ltd., Japan). Using the extracted DNA as a template and the primers of SEQ ID NOs: 1 and 2, quantitative PCR was performed with an Applied Bio systems 7500 Fast & 7500 Real-Time PCR System (Thermo Fisher) to measure the bacterial count of Bifidobacterium bacteria. The PCR conditions were 40 cycles of denaturation at 95°C for 5 seconds, annealing at 55°C for 20 seconds, and extension at 72°C for 30 seconds.

[0072]

Table 2-1

[0073]

Table 2-2

[0074] (4) Results Table 3 shows the bacterial count of Bifidobacterium bacteria after 7-hour culture measured by quantitative PCR. The bacterial count is shown as a relative value when the bacterial count in the non-tocopherol-added group is set to 1. The growth of Bifidobacterium bacteria was further promoted by culturing both 2'-fucosyllactose and 6'-sialyllactose in combination with tocopherol.

[0075]

Table 3-1

[0076]

Table 3-2

[0077]

Table 3-3

[0078]

Table 3-4

Claims

[Claim 1] A nutritional composition comprising oligosaccharides and tocopherol, The nutritional composition (excluding breast milk) wherein the tocopherol comprises one or more tocopherols selected from the group consisting of β-tocopherol, γ-tocopherol and δ-tocopherol.

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