Composition for promoting oligosaccharide assimilation of Bifidobacterium breve

Abstract

The present invention addresses the problem of providing a feature for promoting utilization of an oligosaccharide having a lacto-N-neotetraose skeleton in Bifidobacterium breve. The present invention provides a composition for promoting utilization of an oligosaccharide having a lacto-N-neotetraose skeleton in Bifidobacterium breve, the composition containing one or more saccharides selected from monosaccharides and disaccharides.

Description

[Technical Field]

[0001] The present invention relates to a composition for promoting the assimilation of oligosaccharides by Bifidobacterium breve (B. breve), etc. [Background technology]

[0002] Human milk oligosaccharides (HMOs) are present in human breast milk at a concentration of 10-20 g / L and are said to be a mixture of more than 130 types of oligosaccharides. These oligosaccharides have structures in which fucose and sialic acid are added to 13 types of core structures. Human breast milk has a high proportion of fucosylated neutral sugars, and contains large amounts of 2'-fucosyl lactose (hereinafter also called "2'-FL"), lacto-N-fucopentaose I (hereinafter also called "LNFPI"), lacto-N-difucohexaose I (hereinafter also called "LNDFP I"), and lacto-N-tetraose (hereinafter also called "LNT"). These four types alone account for 1 / 3 to 1 / 4 of all human milk oligosaccharides.

[0003] Oligosaccharides containing lacto-N-biose are called type I, and oligosaccharides containing N-acetyllactosamine (Gal(β1-4)G1cNAc) are called type II. In human breast milk, type I LNT and LNFP1 are present in higher amounts than type II lacto-N-neotetraose (hereinafter also called "LNnT") and lacto-N-fucopentaose III (hereinafter also called "LNFPIII"). The coexistence of type I and type II, and the preference for type I, are distinctive features of human milk oligosaccharides that distinguish them from other types of milk oligosaccharides (Non-Patent Literature 1 and 2). Among these, LNnT, which is type II, is not the main component of HMOs, but reports on safety studies using rats have been published, and it is one of the components that is expected to be put into practical use (Non-Patent Literature 3).

[0004] Incidentally, some Bifidobacterium species commonly found in infants include Bifidobacterium longum subspecies longum, Bifidobacterium longum subspecies infantis, Bifidobacterium breve, and Bifidobacterium bifidum. Of these four species, Bifidobacterium longum subspecies infantis and Bifidobacterium bifidum have been reported to be able to utilize HMOs as a sole carbon source.

[0005] Thus, while some bacteria lack or have low ability to utilize HMOs, many such bacteria can still be used as beneficial bacteria. Therefore, even probiotics that lack or have low ability to utilize HMOs can be useful as nutritional compositions if they can grow in the presence of HMOs.

[0006] To address these challenges, a nutritional composition has been proposed containing bacteria belonging to Bifidobacterium bifidum, one or more probiotics with low human milk oligosaccharide assimilation ability, and human milk oligosaccharides (Patent Document 1). This technology allows for the proliferation of probiotics with low human milk oligosaccharide assimilation ability by utilizing lactose accumulated by bacteria belonging to Bifidobacterium bifidum. However, this technology does not utilize the promotion of HMOs assimilation by the bacteria themselves, which have no or low HMOs assimilation ability. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] WO2019 / 189200 [Non-patent literature]

[0008] [Non-Patent Document 1] Japanese Journal of Lactic Acid Bacteria Vol. 22, No. 1 p.15-25, 2011 [Non-Patent Document 2] Milk Science Vol. 56, No. 4 P.155-176, 2008 [Non-Patent Document 3] Food and Chemical Toxicology Vol.62, p.528-537, 2013 [Overview of the project] [Problems that the invention aims to solve]

[0009] In light of these circumstances, the present invention aims to provide a technology for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton, such as lacto-N-neotetraose (LNnT) of Bifidobacterium breve. [Means for solving the problem]

[0010] The inventors diligently conducted research with the aim of developing a technology to grow probiotics that lack or have low HMOs assimilation ability in the presence of HMOs. In the process, the inventors found that the assimilation ability of Bifidobacterium breve to LNnT is enhanced by coexisting LNnT with one or more sugars selected from monosaccharides and disaccharides.

[0011] In other words, the gist of this invention is as follows: [1] A composition for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve, containing one or more sugars selected from monosaccharides and disaccharides. [2] The composition according to [1], wherein the monosaccharide is one or two selected from glucose and galactose, and the disaccharide is lactose. [3] The composition according to [1] or [2], wherein the Bifidobacterium breve is Bifidobacterium breve NITE BP-02622 or Bifidobacterium breve FERM BP-11175. [4] The composition according to any one of [1] to [3], further containing an oligosaccharide having a lacto-N-neotetraose skeleton. [5] The composition according to any one of [1] to [4], which is a food composition. [6] The composition according to any one of [1] to [5], which is a prepared milk. [7] The composition according to any one of [1] to [4], which is a medium composition. [8] A method for promoting the assimilation of an oligosaccharide having a lacto-N-neotetraose skeleton by Bifidobacterium breve, comprising: coexisting an oligosaccharide having a lacto-N-neotetraose skeleton with one or more sugars selected from monosaccharides and disaccharides. [9] The method according to [8], wherein the monosaccharide is one or two selected from glucose and galactose, and the disaccharide is lactose.

[10] A composition containing one or more sugars selected from monosaccharides and disaccharides, an oligosaccharide having a lacto-N-neotetraose skeleton, and Bifidobacterium breve.

[11] The composition according to

[10] , wherein the monosaccharide is one or two selected from glucose and galactose, and the disaccharide is lactose.

[12] The composition according to

[10] or

[11] , wherein the oligosaccharide having a lacto-N-neotetraose skeleton is lacto-N-neotetraose.

[13] The composition according to any one of

[10] to

[12] , wherein the Bifidobacterium breve is Bifidobacterium breve NITE BP-02622 or Bifidobacterium breve FERM BP-11175.

[0012] Furthermore, the present invention may also employ the following configuration.

[14] A method for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve, comprising administering to one or more sugars selected from monosaccharides and disaccharides.

[15] Use of one or more sugars selected from monosaccharides and disaccharides to promote the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve.

[16] Use of an oligosaccharide having a lacto-N-neotetraose skeleton in the production of any of the compositions described in [1] to [7].

[17] A method for producing the composition according to any one of [1] to [7], comprising formulating one or more sugars selected from monosaccharides and disaccharides. [Effects of the Invention]

[0013] The present invention provides a technology for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton, such as lacto-N-neotetraose of Bifidobacterium breve. The present invention makes it possible to provide useful prebiotic products of Bifidobacterium breve. Furthermore, it is possible to provide useful synbiotic products that combine oligosaccharides having a lacto-N-neotetraose skeleton with Bifidobacterium breve. [Brief explanation of the drawing]

[0014] [Figure 1]Figure 1 shows the results of evaluating the effect of monosaccharides or disaccharides on promoting the assimilation of lacto-N-neotetraose in B. breve. Figure 1A shows the effect of lactose on B. breve FERM BP-11175. Figure 1B shows the effect of glucose on B. breve FERM BP-11175. Figure 1C shows the effect of galactose on B. breve FERM BP-11175 (the LnNT results are almost equivalent to the galactose results and are partially overlapped on the graph). Figure 1D shows the effect of lactose on B. breve NITE BP-02622. Figure 1E shows the effect of glucose on B. breve NITE BP-02622. Figure 1F shows the effect of galactose on B. breve NITE BP-02622. [Modes for carrying out the invention]

[0015] Embodiments of the present invention will be described below. However, the present invention is not limited to the following preferred embodiments and can be freely modified within the scope of the present invention. In this specification, when numerical ranges are expressed as "lower limit to upper limit," the upper limit may be "less than or equal to" or "less than," and the lower limit may be "greater than or equal to" or "greater than."

[0016] In this invention, "probiotics" refers to live microorganisms that, when ingested in appropriate amounts, have beneficial effects on the host's health. "Prebiotics" refers to substances that act as "food" for bifidobacteria and lactic acid bacteria, thereby increasing their numbers and performing beneficial functions. In this invention, probiotics and prebiotics are used in combination as "synbiotics." It is preferable that synbiotics be used as a preparation (intestinal bacteria preparation) made by mixing the two.

[0017] <Inventive Composition> One aspect of the present invention relates to a composition for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve, which contains one or more sugars selected from monosaccharides and disaccharides (hereinafter sometimes referred to as "the composition of the present invention"). The composition of the present invention can be in the form of a food composition, a pharmaceutical composition, or a culture medium composition, etc.

[0018] ≪Sugar≫ The composition of the present invention contains one or more sugars selected from monosaccharides and disaccharides. The monosaccharides and disaccharides that can be used in the composition of the present invention can be freely selected from one or more monosaccharides and disaccharides that can be used in fields such as medicine and food. Specifically, examples include monosaccharides such as glucose, fructose, galactose, rhamnose, and fucose; and disaccharides such as lactose, sucrose, maltose, and trehalose. Preferably, glucose, galactose, and lactose are used, and more preferably, glucose and lactose are used. As monosaccharides and disaccharides, commercially available products may be used, or those that are manufactured as appropriate may be used.

[0019] The content of monosaccharides and disaccharides in the total composition of the present invention is not particularly limited as long as it has the assimilation-promoting effect of the present invention, but may be 0.01% by mass or more, 0.1% by mass or more, 0.2% by mass or more, 1% by mass or more, 2% by mass or more, 5% by mass or more, and may be 100% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less, 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, 20% by mass or less, or 10% by mass or less. Furthermore, the content of monosaccharides and disaccharides in the whole composition may be 0.01% to 100% by mass, 0.1% to 90% by mass, 0.2% to 80% by mass, 1% to 80% by mass, 2% to 70% by mass, or 5% to 60% by mass.

[0020] ≪Bifidobacterium breve≫ The Bifidobacterium breve targeted by the composition of the present invention can be any one or more known or unknown bacteria belonging to Bifidobacterium breve, as long as the effects of the present invention are not impaired. In other words, any bacteria that can promote the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton with one or more sugars selected from monosaccharides and disaccharides can be targeted. The Bifidobacterium breve targeted by the composition of the present invention can be any bacteria that have no ability to assimilate oligosaccharides having a lacto-N-neotetraose skeleton, or have low ability to assimilate them, or have normal assimilation, but preferably bacteria that have no ability to assimilate oligosaccharides having a lacto-N-neotetraose skeleton, or low ability to assimilate them.

[0021] In the present invention, "assimilation of oligosaccharides having a lacto-N-neotetraose skeleton" means "metabolism of oligosaccharides having a lacto-N-neotetraose skeleton." In the present invention, "promotion of assimilation of oligosaccharides having a lacto-N-neotetraose skeleton" means that, when any bacterial strain is cultured under the same conditions, the ability to assimilate oligosaccharides having a lacto-N-neotetraose skeleton is enhanced when the sugar source is monosaccharides or disaccharides and oligosaccharides having a lacto-N-neotetraose skeleton compared to when the sugar source is oligosaccharides having a lacto-N-neotetraose skeleton alone.

[0022] The ability of any bacterial strain to utilize oligosaccharides containing a lacto-N-neotetraose skeleton can be confirmed by known methods, such as by measuring the turbidity, specific growth rate, and sugar consumption rate of the microorganism.

[0023] In the present invention, "promotion of assimilation of oligosaccharides having a lacto-N-neotetraose skeleton" may mean the promotion of growth of Bifidobacterium breve. Specifically, with respect to a culture cultured in a medium containing monosaccharides or disaccharides and oligosaccharides having a lacto-N-neotetraose skeleton as sugar sources, the turbidity (e.g., OD600 value) at a certain point in time may be greater than 1 times the sum of (a) the turbidity (e.g., OD600 value) when the sugar source is monosaccharides or disaccharides only and when the sugar source is oligosaccharides having a lacto-N-neotetraose skeleton, or (b) the turbidity (e.g., OD600 value) when the sugar source is oligosaccharides having a lacto-N-neotetraose skeleton, and may be 1.1 times or more, 1.2 times or more, 1.3 times or more, 1.4 times or more, 1.5 times or more, 2 times or more, 5 times or more, or 10 times or more.

[0024] In the present invention, "promotion of assimilation of oligosaccharides having a lacto-N-neotetraose skeleton" may also mean an increase in the specific growth rate of Bifidobacterium breve (the specific growth rate of microorganisms is defined as the increase in cells per unit time). For example, when oligosaccharides having a lacto-N-neotetraose skeleton are used in combination with one or more sugars selected from monosaccharides and disaccharides as a sugar source, the specific growth rate increases to more than 100%, 200% or more, 500% or more, 1000% or more, 5000% or more, or 6000% or more compared to (a) the sum of the specific growth rates when the sugar source is monosaccharides or disaccharides only and when the sugar source is oligosaccharides having a lacto-N-neotetraose skeleton only, or (b) the specific growth rate when the sugar source contains only oligosaccharides having a lacto-N-neotetraose skeleton.

[0025] In the present invention, "promotion of assimilation of oligosaccharides having a lacto-N-neotetraose skeleton" may mean an increase in the sugar consumption rate of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve. For example, there are no special limitations on the method for measuring the sugar consumption rate, and it may be done by direct or indirect methods. A direct method may be, for example, measuring the sugar concentration in the culture medium using a biosensor. An indirect method may be, for example, measuring the sugar concentration in the culture medium using a refractometer such as a refractometer Brix meter, a vibrating densimeter, a viscometer, etc.

[0026] While not limited to Bifidobacterium breve, examples include Bifidobacterium breve NITE BP-02622, Bifidobacterium breve FERM BP-11175, and Bifidobacterium breve JCM1192.

[0027] The composition of the present invention may also contain Bifidobacterium breve in combination with one or more sugars selected from monosaccharides and disaccharides as active ingredients. That is, "promotion of assimilation of oligosaccharides having a lacto-N-neotetraose skeleton" is not limited to cases where the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve inherently present in the target recipient of the one or more sugars selected from monosaccharides and disaccharides is promoted, but may also include cases where the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve administered to the target recipient together with one or more sugars selected from monosaccharides and disaccharides is promoted.

[0028] Bifidobacterium breve JCM1192 is available from JCM. "JCM" is an abbreviation for the Japan Collection of Microorganisms, BioResource Research Center, RIKEN. Its address is: Postal code: 305-0074, Address: 3-1-1 Takanodai, Tsukuba City, Ibaraki Prefecture, Japan, Japan. In this specification, strains assigned accession numbers beginning with "JCM" are deposited with JCM.

[0029] Bifidobacterium breve NITE BP-02622 can be obtained from NPMD. "NPMD" is an abbreviation for the Patent Microbial Depository Center of the National Institute of Technology and Evaluation, located at Room 122, 2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture 292-0818, Japan. In this specification, strains with accession numbers beginning with "NITE" are deposited with NPMD.

[0030] Bifidobacterium breve FERM BP-11175 is available from NPMD. In 2012, the status of international depositary was transferred from the National Institute of Advanced Industrial Science and Technology (IPOD) to the National Institute of Technology and Evaluation (NITE), and patent microbial depositary operations were centralized under NPMD. In this specification, strains assigned accession numbers beginning with "FERM" were deposited with IPOD and subsequently their management was transferred to NPMD.

[0031] Furthermore, the strains identified by the strain names exemplified above are not limited to the actual strains deposited or registered with the designated institution under those strain names (hereinafter, for convenience of explanation, also referred to as "deposited strains"), but also include strains that are substantially equivalent to them (also referred to as "derived strains" or "inducible strains"). In other words, for example, "Bifidobacterium breve NITE BP-02622" is not limited to the actual strains deposited with the depositary institution under the deposit number Bifidobacterium breve NITE BP-02622, but also includes strains that are substantially equivalent to them. A plant that is substantially equivalent to the deposited plant mentioned above may, for example, be a derivative plant that has the deposited plant as its parent plant. Examples of derivative plants include plants bred from the deposited plant and plants that arose naturally from the deposited plant.

[0032] Examples of stocks that are essentially equivalent or derived from the following stocks include the following: (1) Strains determined to be the same strain by RAPD (Randomly Amplified Polymorphic DNA) method and PFGE (Pulsed-field gel electrophoresis) method (as described on page 43 of Probiotics in food / Health and nutritional properties and guidelines for evaluation, page 85) (2) A strain that possesses only genes originating from the deposited strain, does not have any foreign genes, and has a DNA identity of 95% or more. (3) Strains bred from the strain in question (including genetically modified strains, spontaneous mutations, and strains having the same traits)

[0033] As for Bifidobacterium breve, commercially available products may be used, or products that are manufactured and obtained as appropriate may be used. As a commercially available product of Bifidobacterium breve NITE BP-02622, for example, Bifidobacterium breve M-16V manufactured by Morinaga Milk Industry Co., Ltd. can be used. Furthermore, the bacterial cells of Bifidobacterium breve can be easily obtained by culturing Bifidobacterium breve using known methods.

[0034] The composition of the present invention may also contain Bifidobacterium breve in combination with one or more sugars selected from monosaccharides and disaccharides as active ingredients. In this case, the content of Bifidobacterium breve in the composition of the present invention is not particularly limited as long as it has the assimilation-promoting effect of the present invention, but for example, 10 3 CFU / g or mL or more, 10 4 CFU / g or mL or more, 10 5 CFU / g or mL or more, or 106 It may be 10 CFU / g or mL or more, 10 12 CFU / g or mL or less, 10 11 CFU / g or mL or less, or 10 10 CFU / g or mL or less. The content of Bifidobacterium breve in the composition of the present invention is 10 3 CFU / g or mL to 10 12 CFU / g or mL, and may be 10 4 CFU / g or mL to 10 11 CFU / g or mL, and may be​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​In the present invention, the oligosaccharide having a lacto-N-neotetraose skeleton may be one or more known or unknown oligosaccharides having a lacto-N-neotetraose skeleton, as long as the effects of the present invention are not impaired. That is, by using one or more sugars selected from monosaccharides and disaccharides in combination, it may be an oligosaccharide that can promote the assimilation ability of Bifidobacterium breve. Specifically, examples include lacto-N-neotetraose and sialyllacto-N-neotetraosec.

[0037] The composition according to the present invention may also contain, in combination with one or more sugars selected from monosaccharides and disaccharides as active ingredients, an oligosaccharide having a lacto-N-neotetraose skeleton.

[0038] The content of oligosaccharides having a lacto-N-neotetraose skeleton in the total composition of the present invention is not particularly limited as long as it has the assimilation-promoting effect of the present invention, but may be 0.001% by mass or more, 0.01% by mass or more, 0.1% by mass or more, or 1% by mass or more, and may be 50% by mass or less, 40% by mass or less, 30% by mass or less, 20% by mass or less, 10% by mass or less, 5% by mass or less, or 1% by mass or less. The content of oligosaccharides having a lacto-N-neotetraose skeleton in the total composition of the present invention may be 0.001% to 50% by mass, 0.01% to 40% by mass, 0.1% to 30% by mass, or 1% to 20% by mass.

[0039] The embodiments of the composition of the present invention are not particularly limited as long as they demonstrate the effects of the present invention, and specifically include embodiments such as food compositions, pharmaceutical compositions, or culture medium compositions. The composition of the present invention may be provided in any form such as liquid, paste, gel-like solid, or powder. The content of one or more sugars selected from monosaccharides and disaccharides, the content of Bifidobacterium breve, and the content of LNnT in the food composition, pharmaceutical composition, and culture medium composition of the present invention can be appropriately referenced from the content described in the composition of the present invention.

[0040] As described above, the composition of the present invention may also contain Bifidobacterium breve in combination with one or more sugars selected from monosaccharides and disaccharides, which are active ingredients. Furthermore, the composition of the present invention may also contain, in combination with one or more sugars selected from monosaccharides and disaccharides as active ingredients, an oligosaccharide having a lacto-N-neotetraose skeleton as a prebiotic. Furthermore, one or more known or unknown HMOs other than oligosaccharides having a lacto-N-neotetraose skeleton can be freely selected and used. Among these, 2'-fucosyl lactose, lacto-N-fucopentaose I, lacto-N-difucohexaose I, and lacto-N-tetraose are preferably used. Furthermore, in the present invention, prebiotics other than HMOs may also be used. The prebiotics are not particularly limited as long as they can better exert the effects of the present invention when ingested together with the composition of the present invention. For example, lactulose, raffinose, galactooligosaccharides, fructooligosaccharides, soybean oligosaccharides, lactulose oligosaccharides, xylooligosaccharides, isomaltoligosaccharides, coffee bean mannooligosaccharides, gluconic acid, polydextrose, inulin, etc. are preferred, and among these, lactulose, raffinose, galactooligosaccharides, etc. are more preferred, and lactulose, raffinose, and galactooligosaccharides are even more preferred.

[0041] The above-mentioned composition of the present invention can be used, for example, as a synbiotic combining probiotics and prebiotics in the form of a food composition or a pharmaceutical composition. When the composition of the present invention is a food composition or pharmaceutical composition to be administered to a target, the monosaccharide and disaccharide content in the composition of the present invention can be determined by applying mutatis mutandis the description of the dosage of monosaccharides and disaccharides in the method of the present invention. Administering the composition of the present invention to a target can be expected to promote the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve. In other words, the proliferation of probiotics in the intestines improves the gut microbiota. By improving the gut microbiota, diseases caused by the gut microbiota may be improved, prevented, or become treatable. The use of this embodiment may be for therapeutic purposes or for non-therapeutic purposes. "Non-therapeutic purposes" is a concept that does not include medical procedures, that is, treatments performed on the human body. Examples include health promotion and cosmetic procedures. "Improvement" means a rise in the disease, symptoms, or condition; prevention or delay of the worsening of a disease, symptoms, or condition; or a reversal, prevention, or delay of the progression of a disease or condition. "Prevention" means preventing or delaying the onset of a disease or symptom in the target area, or reducing the risk of the disease or symptom in the target area.

[0042] The target population for administration of the composition of the present invention is not particularly limited, as long as the desired effects, such as assimilation-promoting effects, can be obtained, but examples include mammals. Examples of mammals include humans and pets. Examples of pets include dogs and cats. Humans are particularly mentioned as mammals. The composition of the present invention may be administered to any human who desires the desired effects, such as proliferation-promoting effects. Humans may be of any age, such as infants, toddlers, children, adults, middle-aged and elderly people, etc. "Infants" refers to children under one year old. "Toddlers" refers to children from one year old until they start elementary school. Infants and toddlers are preferred targets for administration of the composition of the present invention.

[0043] When the composition of the present invention is used for non-therapeutic purposes, it may be administered to healthy individuals. Healthy individuals may mean those who do not suffer from diseases caused by the intestinal microbiota, those who are not in an unhealthy state due to a disruption of the intestinal microbiota, or those who do not have abnormalities in their immune function. When the composition of the present invention is used for non-therapeutic purposes, it may be used to further improve the healthy intestinal microbiota possessed by healthy individuals, and in particular, it may be used to promote the growth of Bifidobacterium breve in the intestines of healthy individuals. When the composition of the present invention is used for non-therapeutic purposes, it may be possible to prevent diseases caused by intestinal bacteria in healthy individuals. Diseases caused by the intestinal microbiota include inflammatory bowel disease, ulcerative colitis, irritable bowel disease, colon cancer, diabetes mellitus, and arteriosclerosis. Furthermore, when the composition of the present invention is used for non-therapeutic purposes, it further enhances the normal immune function in healthy individuals, thereby making it possible to prevent allergic symptoms such as hay fever, asthma, and atopic dermatitis in healthy individuals.

[0044] When the composition of the present invention is used for therapeutic purposes, it may be administered to non-healthy individuals. Non-healthy individuals include those who exhibit symptoms for which administration of Bifidobacterium breve is effective in prevention, improvement, and / or treatment, such as those suffering from diseases caused by the gut microbiota or those with immune system abnormalities. Diseases caused by the gut microbiota include inflammatory bowel disease, ulcerative colitis, irritable bowel syndrome, colon cancer, diabetes mellitus, and arteriosclerosis. Immune system abnormalities include allergic symptoms such as hay fever, asthma, and atopic dermatitis. When the compositions of the present invention are used for therapeutic purposes, it may be possible to improve, prevent, and treat diseases caused by the gut microbiota of unhealthy individuals, or abnormalities in immune function. When the composition of the present invention is used for therapeutic purposes, it can be used to improve the intestinal microbiota of non-healthy individuals, and in particular, to promote the growth of Bifidobacterium breve in the intestines of non-healthy individuals.

[0045] <Inventive Food Composition> One aspect of the present invention relates to a food composition, which is the composition of the present invention (hereinafter sometimes referred to as "the food composition of the present invention"). The food composition according to the present invention can be added as is or in combination with formulation components to known foods and beverages as an additive, used as a supplement, or mixed into the raw materials of foods and beverages and used as a food or beverage. As formulation components, refer to the formulation carriers described in <Pharmaceutical Compositions> below. The method for producing the food composition of the present invention is not particularly limited. The food composition of the present invention can be produced, for example, using the same raw materials as ordinary foods and beverages and using the same methods as ordinary foods and beverages, except for the addition of one or more sugars selected from monosaccharides and disaccharides.

[0046] Foods and beverages using the food composition according to the present invention can be in any form, such as liquid, paste, solid, or powder, and include tablets, liquid foods, and other products such as wheat flour products, instant foods, processed agricultural products, processed marine products, processed livestock products, milk and dairy products, oils and fats, basic seasonings, compound seasonings and foods, frozen foods, confectionery, beverages, and other commercially available products.

[0047] Examples of the aforementioned wheat flour products include bread, macaroni, spaghetti, noodles, cake mix, fried chicken batter, and breadcrumbs. Examples of the aforementioned instant foods include instant noodles, cup noodles, retort / prepared foods, canned prepared foods, microwaveable foods, instant soups / stews, instant miso soup / clear soup, canned soups, freeze-dried foods, and other instant foods. Examples of the aforementioned processed agricultural products include canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, and cereals (processed grain products). Examples of the aforementioned processed seafood products include canned seafood, fish ham and sausage, processed seafood products, seafood delicacies, and preserved seafood products. Examples of the aforementioned processed livestock products include canned and pasteurized livestock products, and meat ham and sausages. Examples of the milk and dairy products mentioned above include fermented milk, processed milk, milk beverages, yogurts, lactic acid bacteria beverages, cheese, ice cream, prepared milk powder, cream, and other dairy products. Examples of the aforementioned fats and oils include butter, margarine, and vegetable oil. Examples of the basic seasonings include soy sauce, miso, sauces, tomato-based seasonings, mirin, and vinegars, while examples of the compound seasonings and foods include cooking mixes, curry bases, sauces, dressings, noodle soup bases, spices, and other compound seasonings. Examples of the aforementioned frozen foods include frozen raw ingredients, partially cooked frozen foods, and pre-cooked frozen foods. Examples of the aforementioned confectionery include caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice crackers, bean sweets, dessert sweets, and other confectionery. Examples of the aforementioned beverages include carbonated drinks, natural fruit juices, fruit juice beverages, fruit juice-containing soft drinks, fruit pulp beverages, fruit pulp beverages, vegetable-based beverages, soy milk, soy milk beverages, coffee beverages, tea beverages, powdered beverages, concentrated beverages, sports drinks, nutritional beverages, alcoholic beverages, and other beverages for enjoyment. Other commercially available foods besides those mentioned above include, for example, baby food, furikake (rice seasoning), and tea-soaked seaweed.

[0048] Furthermore, the food compositions defined in this invention can also be provided and sold as food and beverages with specific uses (especially health uses) or functions indicated. The act of "displaying" includes all acts that inform consumers of the aforementioned use, and any expression that can evoke or infer the aforementioned use falls under the category of "displaying" according to the present invention, regardless of the purpose of the display, the content of the display, or the object or medium to which it is displayed.

[0049] Furthermore, the "display" should preferably be made in a way that allows consumers to directly recognize the above-mentioned use. Specifically, this includes acts such as transferring, delivering, displaying for transfer or delivery, or importing food and beverage products or product packaging on which the above-mentioned use is described; displaying or distributing advertisements, price lists, or transaction documents related to products that describe the above-mentioned use; or providing information containing such information by electromagnetic means (such as the Internet).

[0050] On the other hand, the content of the display is preferably a display approved by the government or other administrative body (for example, a display approved based on various systems established by the government and made in accordance with such approval). Furthermore, it is preferable to attach such display content to packaging, containers, catalogs, brochures, point-of-sale (POP) displays and other promotional materials used at sales sites, and other documents.

[0051] Furthermore, "labeling" also includes labels such as health foods, functional foods, foods for the sick, enteral nutrition foods, foods for special dietary uses, health functional foods, foods for specified health uses, foods with functional claims, and foods with nutritional function. In particular, labels approved by the Consumer Affairs Agency include, for example, labels approved under the Foods for Specified Health Uses system, the Foods with Functional Claims system, and similar systems. More specifically, these include labels as Foods for Specified Health Uses, labels as conditionally approved Foods for Specified Health Uses, labels as Foods with Functional Claims, labels indicating an effect on the structure or function of the body, and labels indicating a reduction in disease risk. Typical examples of these include labels as Foods for Specified Health Uses (especially labels indicating health uses) as defined in the Enforcement Regulations of the Health Promotion Act (Ministry of Health, Labour and Welfare Ordinance No. 86 of April 30, 2003), labels as Foods with Functional Claims as defined in the Food Labeling Act (Act No. 70 of 2013), and similar labels. Specific examples of labels include, "Bifidobacterium breve increases in the stomach," and "Assimilates LNnT, a human milk oligosaccharide."

[0052] The food composition of the present invention can be administered to any subject, including healthy and unhealthy individuals. However, when it is presented as a food or beverage with a specific use (especially a health use) or function indicated, it is used for the non-therapeutic purposes described above.

[0053] <Made-to-feed milk> The food composition according to the present invention can be suitably used in food and beverages, particularly in prepared milk. Examples of prepared milk include prepared powdered milk and prepared liquid milk. Examples of formulated milk include infant formula, follow-up formula, formula for low birth weight infants, formula for children, formula for adults, formula for the elderly, formula for allergies, formula for lactose intolerance, and formula for congenital metabolic disorders. It is particularly preferable to use it for infant formula, follow-up formula, formula for low birth weight infants, and formula for children. Formula for infants and young children refers to infant formula for infants aged 0-12 months, follow-up formula for infants aged 6-9 months and older and young children (up to 3 years old), formula for low birth weight infants (newborns weighing less than 2500g at birth), and various therapeutic formulas used to treat children with pathological conditions such as milk allergy or lactose intolerance.

[0054] Furthermore, this composition can be applied to health functional foods and foods for sick people. The health functional food system was established based on domestic and international trends and consistency with the conventional system for specified health foods, and applies not only to ordinary foods but also to foods in the form of tablets, capsules, etc. It consists of two types: specified health foods (individually approved type) and nutrient function foods (standard type).

[0055] The method for producing the prepared milk can be any combination of one or more known production methods, as long as the effects of the present invention are not impaired. Specifically, for example, the food composition and various additives according to the present invention can be mixed with raw milk, and the product can be produced through processes such as homogenization, sterilization, concentration, drying, granulation, and sieving.

[0056] The prepared milk of the present invention can be manufactured, for example, by the following method.

[0057] In other words, the present invention provides a method for producing powdered milk by mixing one or more sugars selected from monosaccharides and disaccharides (and optionally, further containing a bacterial cell powder containing Bifidobacterium breve and an oligosaccharide having a lacto-N-neotetraose skeleton) with milk components to obtain powdered milk. Specifically, the present invention provides a method for producing powdered milk, which includes the following step (A): (A) A step of mixing one or more sugars selected from monosaccharides and disaccharides (and optionally, cell powder containing Bifidobacterium breve and oligosaccharides having a lacto-N-neotetraose skeleton) and milk components to obtain powdered milk.

[0058] Furthermore, the food composition of the present invention may specifically be, for example, a supplement for infants. A supplement for infants can be manufactured, for example, by the following method. Specifically, the present invention provides a method for manufacturing supplements for infants and young children, comprising the following steps (A) and (B): (A) A step of mixing one or more sugars selected from monosaccharides and disaccharides (and optionally, further containing bacterial cell powder containing Bifidobacterium breve and oligosaccharides having a lacto-N-neotetraose skeleton) and excipients to obtain a mixture; (B) Step of compressing the mixture into tablets.

[0059] In any of the above manufacturing methods, other components besides those mentioned in the above steps may be used in combination as appropriate.

[0060] The method of adding the food composition according to the present invention is not particularly limited as long as it does not impair the effects of the present invention. For example, in addition to adding the food composition according to the present invention in powder or liquid form during mixing as described above, it can also be added after pulverization and mixed as a powder during production.

[0061] The prepared milk of the present invention can be administered to any subject, including healthy individuals and those who are not healthy. When the prepared milk of the present invention is administered to healthy individuals, it can be used as infant formula, follow-up formula, formula for low birth weight infants, formula for children, formula for adults, and formula for the elderly, and can be used for the non-therapeutic purposes described above.

[0062] When the prepared milk of the present invention is administered to non-healthy individuals, it can be used as a prepared milk for allergies, lactose intolerance, or congenital metabolic disorders, and can be used for the above-mentioned therapeutic purposes.

[0063] <Inventive Culture Medium Composition> One aspect of the present invention relates to a culture medium composition, which is a composition of the present invention (hereinafter sometimes referred to as "the culture medium composition of the present invention"). The composition according to the present invention can be added as is or in combination with formulation components to known culture media as an additive, or it can be mixed into the raw materials of a culture media and used as a culture media composition. The culture media composition may be a culture media composition for Bifidobacterium breve. As formulation components, refer to the formulation carriers described in <Pharmaceutical Compositions> below. The method for producing the culture media composition of the present invention is not particularly limited. The culture media composition of the present invention can be produced, for example, using the same raw materials as a conventional culture media and in the same manner as a conventional culture media, except for the addition of one or more sugars selected from monosaccharides and disaccharides.

[0064] When the composition according to the present invention is incorporated into a culture medium, the raw materials for the culture medium include, for example, a carbon source, a nitrogen source, inorganic salts, organic components, and milk components. The raw materials for the culture medium may consist of one component, or a combination of two or more components. Examples of carbon sources include sugars other than the active ingredient of the composition according to the present invention, such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch, starch hydrolysates, and molasses. Examples of nitrogen sources include ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, as well as nitrates. Examples of inorganic salts include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate. Examples of organic components include peptone, soybean flour, defatted soybean meal, meat extract, and yeast extract. Examples of milk components include milk protein. Furthermore, as a culture medium, for example, one or more sugars selected from monosaccharides and disaccharides can be added to a medium commonly used for culturing Bifidobacterium bacteria such as Bifidobacterium breve. Examples of media 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).

[0065] <Pharmaceutical composition> The composition according to the present invention can be used as a pharmaceutical or quasi-drug (hereinafter also referred to as "pharmaceutical composition") as is, or it can be used as a pharmaceutical composition by mixing it with the raw materials of a pharmaceutical or quasi-drug. The pharmaceutical composition can be manufactured, for example, using the same raw materials as a conventional pharmaceutical composition and in the same manner as a conventional pharmaceutical composition, except for the addition of one or more sugars selected from monosaccharides and disaccharides.

[0066] When the composition according to the present invention is used as a pharmaceutical composition, the pharmaceutical composition can be formulated into a desired dosage form as appropriate, depending on the method of administration, such as oral or parenteral administration. The dosage form is not particularly limited, but in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets, lozenges, and capsules; or liquid preparations such as solutions, syrups, suspensions, and emulsions. In the case of parenteral administration, it can be formulated into suppositories, sprays, inhalants, ointments, patches, injections, etc. In the present invention, formulation into an oral dosage form is preferred. The formulation can be carried out using known methods as appropriate, depending on the dosage form.

[0067] During formulation, a formulation carrier may be added as appropriate. In addition to the active ingredient of the present invention, other ingredients commonly used in formulation, such as excipients, pH adjusters, colorants, and flavoring agents, can be used. Furthermore, ingredients that have preventive, ameliorative, and / or therapeutic effects on known or potentially discovered diseases and symptoms may be used in combination as appropriate for the purpose.

[0068] As the aforementioned formulation carrier, various organic or inorganic carriers can be used depending on the dosage form. Examples of carriers in the case of solid formulations include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, and odor-masking agents.

[0069] Examples of the 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, carboxymethylcellulose, and carboxymethylcellulose calcium; 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.

[0070] Examples of the binders include, in addition to the above-mentioned excipients, gelatin; polyvinylpyrrolidone; macrogol, and the like.

[0071] Examples of the disintegrant include, in addition to the above-mentioned excipients, chemically modified starches or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and cross-linked polyvinylpyrrolidone.

[0072] Examples of the aforementioned lubricants include talc; stearic acid; metal stearate salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as beegum and gayl wax; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylate salts 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 anhydrous silicic acid and silicic acid hydrate; and starch derivatives.

[0073] Examples of the aforementioned stabilizers include para-hydroxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

[0074] Examples of the aforementioned flavoring and odor-correcting agents include sweeteners, acidulants, and flavorings. In the case of liquid formulations for oral administration, examples of carriers used include solvents such as water, and flavoring and odor-masking agents.

[0075] The pharmaceutical composition of the present invention is administered to non-healthy individuals and used for the therapeutic purposes described above.

[0076] <Method of the present invention> One aspect of the present invention relates to a method for promoting the assimilation of Bifidobacterium breve oligosaccharides having a lacto-N-neotetraose skeleton, the method comprising coexisting the oligosaccharide having a lacto-N-neotetraose skeleton with one or more sugars selected from monosaccharides and disaccharides (hereinafter sometimes referred to as "the method of the present invention"). In other words, the method of the present invention is a method that includes the step of coexisting an oligosaccharide having a lacto-N-neotetraose skeleton with one or more sugars selected from monosaccharides and disaccharides.

[0077] The present invention may be a method for producing a composition for promoting the assimilation of Bifidobacterium breve oligosaccharides having a lacto-N-neotetraose skeleton, comprising the step of mixing an oligosaccharide having a lacto-N-neotetraose skeleton with one or more sugars selected from monosaccharides and disaccharides.

[0078] The present invention may be a method for culturing Bifidobacterium breve, for example, comprising the step of culturing Bifidobacterium breve in a culture medium containing one or more sugars selected from monosaccharides and disaccharides. The oligosaccharide having a lacto-N-neotetraose skeleton may already be contained in the culture medium and may be made to coexist by adding one or more sugars selected from monosaccharides and disaccharides, or it may coexist as a culture medium composition containing the oligosaccharide having a lacto-N-neotetraose skeleton and one or more sugars selected from monosaccharides and disaccharides.

[0079] The present invention may be a method for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve, for example, by administering one or more sugars selected from monosaccharides and disaccharides to a subject. Note that "administering to a subject" may be synonymous with "ingesting to a subject". The oligosaccharide having a lacto-N-neotetraose skeleton may be coexisted with one or more sugars already present in the subject and selected from monosaccharides and disaccharides by adding them, or it may be coexisted by administering a composition containing the oligosaccharide having a lacto-N-neotetraose skeleton and one or more sugars selected from monosaccharides and disaccharides.

[0080] The dosage of one or more sugars selected from monosaccharides and disaccharides may be, for example, 1 μg / kg body weight / day or more, 2 μg / kg body weight / day or more, 5 μg / kg body weight / day or more, 10 μg / kg body weight / day or more, 20 μg / kg body weight / day or more, 50 μg / kg body weight / day or more, 100 μg / kg body weight / day or more, or 200 μg / kg body weight / day or more, or 10,000 μg / kg body weight / day or less, 1,000 μg / kg body weight / day or less, 500 μg / kg body weight / day or less, 200 μg / kg body weight / day or less, 100 μg / kg body weight / day or less, 50 μg / kg body weight / day or less, 20 μg / kg body weight / day or less, or 10 μg / kg body weight / day or less, or within a range of combinations thereof. The dosage of one or more sugars selected from monosaccharides and disaccharides may specifically be, for example, 1 μg / kg body weight / day to 10,000 μg / kg body weight / day, 2 μg / kg body weight / day to 1,000 μg / kg body weight / day, or 5 μg / kg body weight / day to 200 μg / kg body weight / day.

[0081] The duration of administration of one or more sugars selected from monosaccharides and disaccharides may be, for example, 4 weeks or more, preferably 8 weeks or more, and more preferably 12 weeks or more. One or more sugars selected from monosaccharides and disaccharides may be administered, for example, once a day, or divided into multiple doses per day. Also, one or more sugars selected from monosaccharides and disaccharides may be administered, for example, daily, or once every few days. One or more sugars selected from monosaccharides and disaccharides may be administered daily in particular. The amount of one or more sugars selected from monosaccharides and disaccharides administered at each dose may be constant or not.

[0082] The matters described in the method of the present invention can be applied to other embodiments of the present invention, namely, compositions, food compositions, prepared milks, and pharmaceutical compositions.

[0083] When the method of the present invention is used for non-therapeutic purposes, it can be used in accordance with the non-therapeutic use described above.

[0084] When the method of the present invention is used for therapeutic purposes, it can be used in accordance with the above-described use for therapeutic purposes.

[0085] One aspect of the present invention relates to a composition containing one or more sugars selected from monosaccharides and disaccharides, an oligosaccharide having a lacto-N-neotetraose skeleton, and Bifidobacterium breve.

[0086] The composition of the present invention described above is expected to promote the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve.

[0087] The embodiments of the composition of the present invention are not particularly limited as long as they demonstrate the effects of the present invention. The embodiments of the composition of the present invention can be any appropriate form depending on the intended use of the composition. Specifically, embodiments of the composition of the present invention include food compositions, pharmaceutical compositions, or culture medium compositions. For example, the composition of the present invention can be used as a synbiotic, combining probiotics and prebiotics, in the form of a food composition or pharmaceutical composition.

[0088] Furthermore, the matters described in the above sections on <Composition of the Invention>, <Food Composition of the Invention>, <Prepared Milk of the Invention>, <Culture Medium Composition of the Invention>, <Pharmaceutical Composition>, and <Method of the Invention> all apply to the description of the above-mentioned compositions of the Invention. That is, for example, the matters described in the above sections on <Composition of the Invention>, <Food Composition of the Invention>, <Prepared Milk of the Invention>, <Culture Medium Composition of the Invention>, <Pharmaceutical Composition>, and <Method of the Invention> can be applied to matters such as the components contained in the above-mentioned compositions of the Invention, methods of use, and methods of manufacture.

[0089] When using the composition of the present invention for non-therapeutic purposes, it can be used in accordance with the above-described non-therapeutic use.

[0090] When the composition of the present invention is used for therapeutic purposes, it can be used in accordance with the above-described therapeutic use. [Examples]

[0091] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples unless it exceeds the essence of the invention.

[0092] <Example Test> This study was conducted to evaluate the effect of monosaccharides or disaccharides on promoting the assimilation of lacto-N-neotetraose by Bifidobacterium breve. (Preparation of culture medium) 80 mL of sugar-free MRS medium (Table 1) was prepared, adjusted to pH 6.5, and then sterilized at 121°C for 15 minutes. LNnT solution, glucose solution, lactose solution, and galactose solution (Table 2) were prepared separately and filtered sterilized using a 0.22 μm filter (Merck Millipore). The resulting preparations were added to the sterilized sugar-free MRS medium to achieve final concentrations of 1% (w / v) for LNnT, and 0.2% (w / v) for glucose, lactose, or galactose, respectively, to prepare a total of seven different media (Table 3).

[0093] [Table 1]

[0094] [Table 2]

[0095] [Table 3]

[0096] (Culture experiment) Using multi-well plates (Falcon), Fidobacterium breve FERM BP-11175 and Bifidobacterium breve NITE BP-02622 were pre-cultured in MRS medium (Difco) supplemented with 0.05% cysteine ​​(Kanto Chemical Co., Ltd.). Subsequently, each pre-cultured strain was divided into three sets of 1 × 10⁶ samples in each of the seven prepared media. 9 A 1% inoculation of approximately cfu / ml was performed, and the cultures were incubated at 37°C for 12 hours. Turbidity (OD600nm) was measured every 30 minutes using a tunable absorbance microplate reader (TECAN Sunrise Rainbow Thermo RC-R). The culture medium without bacterial inoculation was used as the baseline. Three cultures were measured three times each, and the average value was calculated.

[0097] (result) Figure 1 shows the growth curves for each bacterial strain in each culture medium. A higher turbidity value compared to the start of cultivation suggests an increase in the number of Bifidobacteria, indicating that they are utilizing sugars. In both Bifidobacterium breve FERM BP-11175 and Bifidobacterium breve NITE BP-02622 strains, when LNnT was used as the sole sugar source, the turbidity values ​​remained largely unchanged or increased only slightly over time. On the other hand, when LNnT was used in combination with a monosaccharide or disaccharide (glucose, galactose, or lactose) as the sugar source, the turbidity values ​​increased significantly with the progression of culture time compared to when LNnT was used alone or when only a monosaccharide or disaccharide was used. It was confirmed that combining LNnT with one of the selected sugars (glucose, galactose, or lactose) enhanced the LNnT assimilation ability of Bifidobacterium breve FERM BP-11175 and Bifidobacterium breve NITE BP-02622, i.e., promoted LnNT assimilation.

[0098] As described above, it has become clear that monosaccharides or disaccharides can promote the assimilation of oligosaccharides containing the lacto-N-neotetraose skeleton of Bifidobacterium breve.

[0099] Examples of the production of the pharmaceutical and food / beverage compositions of the present invention are shown below, but the compositions of this technology are not limited thereto.

[0100] [Manufacturing Example 1] Bifidobacterium breve FERM BP-11175 or Bifidobacterium breve NITE BP-02622 is added to 3 mL of MRS liquid medium and cultured anaerobically at 37°C for 16 hours. The culture solution is then concentrated and freeze-dried to obtain freeze-dried bacterial powder (bacterial powder). The bacterial powder is uniformly mixed with a monosaccharide or disaccharide (glucose, galactose, or lactose) and lacto-N-neotetraose to obtain a composition. 20 g of this composition is dissolved in 200 g of water to obtain a composition containing Bifidobacterium breve, a monosaccharide or disaccharide, and LNnT. Administering this composition to a target is expected to promote the assimilation of LNnT by Bifidobacterium breve in the intestines.

[0101] [Manufacturing Example 2] Bifidobacterium breve FERM BP-11175 or Bifidobacterium breve NITE BP-02622 is added to 3 mL of MRS liquid medium and incubated anaerobically at 37°C for 16 hours. The culture solution is then concentrated and freeze-dried to obtain freeze-dried bacterial powder (bacterial powder). Next, monosaccharides or disaccharides (glucose, galactose, or lactose), lacto-N-neotetraose, and crystalline cellulose are added to a stirring granulator and mixed. Purified water is then added and granulated, and the granules are dried to obtain granules (pharmaceutical composition) containing Bifidobacterium breve, monosaccharides or disaccharides, and LNnT, as well as excipients. Administering these granules to the target population is expected to promote the assimilation of LNnT by Bifidobacterium breve in the intestines.

[0102] [Manufacturing Example 3] The method for producing fermented milk to which Bifidobacterium breve, monosaccharides or disaccharides, and LNnT are added is shown below.

[0103] First, the milk raw materials, along with water and other components as needed, are mixed and preferably homogenized, followed by heat sterilization. Homogenization and heat sterilization can be carried out by conventional methods. Lactic acid bacteria starter is added (inoculated) to the heat-sterilized milk solution, and fermentation is carried out at a predetermined fermentation temperature to obtain a fermented product. Curd is formed by fermentation.

[0104] As a lactic acid bacteria starter, you can use lactic acid bacteria commonly used in yogurt production, such as Lactobacillus bulgaricus, Lactococcus lactis, and Streptococcus thermophilus. Once the pH reaches the target value, the formed curd is crushed by stirring and cooled to below 10°C to obtain the fermented product. Cooling to below 10°C reduces the activity of the lactic acid bacteria and suppresses acid production.

[0105] Next, the fermented product obtained in the fermentation process is heat-treated to obtain a post-heated fermented product (fermented product after heat treatment). By moderately heating the fermented product, the production of acid by lactic acid bacteria in the post-heated fermented product can be suppressed. This suppresses the decrease in pH during subsequent manufacturing processes and / or during storage of concentrated fermented milk containing bifidobacteria, and as a result, the survival rate of bifidobacteria can be improved.

[0106] Next, Bifidobacterium breve FERM BP-11175 or Bifidobacterium breve NITE BP-02622, a monosaccharide or disaccharide (glucose, galactose, or lactose), and lacto-N-neotetraose are added to the post-fermented product obtained in the heat treatment process. The amount of Bifidobacterium breve added is 1 × 10⁻⁶ relative to the post-fermented product. 7 ~1 × 10 11 cfu / ml is preferred, 1 × 10 8 ~1 × 10 10 CFU / ML is more preferable.

[0107] After heating, Bifidobacterium breve, monosaccharides or disaccharides, and LNnT are added to the fermented product, followed by concentration. The concentration process can be carried out using any known concentration method as appropriate. For example, centrifugation or membrane separation can be used. In centrifugation, whey is removed from the concentrate (post-fermented product to which Bifidobacterium breve, monosaccharides or disaccharides, and LNnT have been added), resulting in concentrated fermented milk containing bifidobacteria and prebiotics with a higher solid content.

[0108] By administering the fermented milk obtained as described above, it is expected that the assimilation of LNnT by Bifidobacterium breve in the intestines will be promoted.

[0109] [Manufacturing Example 4] The method for producing a prepared milk powder containing Bifidobacterium breve, a monosaccharide or disaccharide, and LNnT is shown below.

[0110] 10 kg of desalted milk whey protein powder (Mirai Co., Ltd.), 6 kg of milk casein powder (Fonterra Co., Ltd.), 48 kg of lactose (Mirai Co., Ltd.), 920 g of mineral mixture (Tomita Pharmaceutical Co., Ltd.), 32 g of vitamin mixture (Tanabe Pharmaceutical Co., Ltd.), 500 g of lactulose (Morinaga Milk Industry Co., Ltd.), and 500 g of lact-N-neotetraose are dissolved in 300 kg of warm water, and then heated and dissolved at 90°C for 10 minutes. 28 kg of prepared fat (Taiyo Yushi Co., Ltd.) is added and homogenized. After that, sterilization and concentration processes are carried out, and spray drying is performed to prepare approximately 95 kg of prepared milk powder. To this, the bacterial cell powder (1.8 × 10) of Bifidobacterium breve FERM BP-11175 or Bifidobacterium breve NITE BP-02622 dispersed in starch is added. 11 Add 100g of cfu / g (manufactured by Morinaga Milk Industry Co., Ltd.) to prepare approximately 95kg of formula milk containing Bifidobacterium breve, monosaccharides or disaccharides, and LNnT. When the resulting formula milk is dissolved in water to make a formula solution with a total solids content of 14% (w / v), which is the standard formula concentration, the number of Bifidobacteria in the formula solution is 2.7 × 10⁶. 9 It is possible to obtain cfu / 100mL.

[0111] By administering the prepared milk powder obtained as described above, it is expected that the assimilation of LNnT by Bifidobacterium breve will be promoted in the intestines. [Industrial applicability]

[0112] This invention is useful in fields such as food and beverages, health foods, functional foods, supplements, culture media, and pharmaceuticals.

[0113] TIFF0007854503000004.tif232169

Claims

1. A composition for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve, comprising one or more sugars selected from glucose, galactose, and lactose.

2. The composition according to claim 1, wherein the Bifidobacterium breve is Bifidobacterium breve NITE BP-02622 or Bifidobacterium breve FERM BP-11175.

3. The composition according to claim 1 or 2, further comprising an oligosaccharide having a lacto-N-neotetraose skeleton.

4. A food composition according to claim 1 or 2.

5. The composition according to claim 1 or 2, which is a prepared milk.

6. The composition according to claim 1 or 2, which is a culture medium composition.

7. A method for promoting the assimilation of oligosaccharides having a lacto-N-neotetraose skeleton of Bifidobacterium breve, Oligosaccharides having a lacto-N-neotetraose skeleton, A method comprising coexisting with one or more sugars selected from glucose, galactose, and lactose.

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