WO2025116014A1 - Composition for controlling proliferation of subdoligranulum bacteria - Google Patents

Abstract

The present invention addresses the problem of providing a means for controlling the proliferation of Subdoligranulum bacteria. Provided is a composition for controlling the proliferation of Subdoligranulum bacteria, the composition comprising a component selected from between kojibiose and an oligosaccharide having kojibiose as a constituent sugar. In one aspect, the composition comprises galactosyl kojibiose as the component selected from between kojibiose and an oligosaccharide having kojibiose as a constituent sugar.

Description

Composition for controlling the growth of bacteria belonging to the genus Subdoligranulum

The present invention relates to a composition for controlling the growth of bacteria of the genus Subdoligranulum.

　Subdoligranulum bacteria are bacteria that live in the intestines of mammals, including humans. They are known to be less prevalent in children with food allergies and in patients with inflammatory bowel disease (Non-Patent Documents 1 and 2). It has also been shown that administering Subdoligranulum bacteria to food allergy model mice can suppress food allergy symptoms (Non-Patent Document 1), and it is known that Subdoligranulum bacteria show a positive correlation with HDL cholesterol (Non-Patent Document 3) and a negative correlation with insulin resistance, body weight, and C-reactive protein (CRP), whose blood concentration rises rapidly when there is inflammation or tissue destruction (Non-Patent Document 4).

Regarding Subdoligranulum bacteria, Patent Document 1 describes a composition of a produced strain of Subdoligranulum bacteria, and a method for treating a pathogen infection, such as Clostridium difficile infection, by administering the composition to a subject having the pathogen infection. Patent Document 2 describes the use of grape skin or a composition containing the same as a prebiotic to increase intestinal levels and/or activity of butyric acid-producing bacteria, such as Faecalibacterium prausnitzii, Phylogenetic Group I (PHGI), Phylogenetic Group II (PHGII), Roseburia hominis, and Subdoligranulum variabile, in a subject. Furthermore, Patent Document 3 describes a composition and method for inducing and/or expanding CD8+ T cells, the composition comprising a purified bacterial mixture containing two or more bacterial strains of species selected from Fusobacterium ulcerans and Eubacterium limosum, or Bacteroides dorei and Bacteroides uniformis, and the composition may further comprise a bacterium of the genus Subdoligranulum. Patent Document 4 describes a pharmaceutical composition comprising a minimal microbial consortium of culturable species and a pharma- ceutically acceptable carrier, the consortium options being a preparation of a viable and culturable anaerobic enterobacteria strain expressing exopolysaccharide, lipoteichoic acid (LTA), lipopolysaccharide (LPS) or other microbial adjuvant molecules that promote the development of regulatory T cells (Treg). This document describes Subdoligranulum variabile as one of the anaerobic enterobacteria strains.

Furthermore, Patent Document 5 describes a method for treating cancer in a mammalian subject, comprising administering to the subject a therapeutic composition containing an effective amount of an isolated population of bacteria belonging to one or more of the genera Ruminococcus, Gemiger, Faecalibacterium, and Subdoligranurum, or a combination thereof. Patent Document 6 describes an agent for regulating the number of bacteria in the intestine belonging to at least one family or genus selected from the group consisting of the family Ruminococcus, the genus Catenibacterium, the genus Subdoligranurum, the genus Family XIII AD3011 group, and the genus Anaeroturuncus, containing propolis as an active ingredient. This document also describes that the agent for regulating the number of intestinal bacteria can be used for preventing or improving endometrial cancer, preventing or improving polycystic ovary syndrome, preventing or improving kidney disease, preventing or improving endotoxemia or inflammatory markers in type 1 diabetes, etc., by reducing the number of the above-mentioned bacteria in the intestine.

On the other hand, it is known that kojibiose has a growth promoting effect on bifidobacteria (e.g., Non-Patent Document 5). Furthermore, Patent Document 7 describes an in vivo lipid regulator containing one or more active ingredients selected from kojibiose, kojitriose, kojibiosyl glucoside, kojitetraose, and kojitriosyl glucoside. Furthermore, Patent Document 8 describes a dosage form containing a composition containing a microbiome regulator, and sugar or sugar alcohol is listed as one of the ingredients that can be contained in the agent, with kojibiose being listed as an example, and the genus Subdoligranulum is listed as one of the endogenous symbiotic microbial taxa or exogenously administered probiotic bacterial taxa whose growth is regulated by the microbiome regulator. It is also described that the agent is used to treat subjects with intestinal dysbiosis, and examples of intestinal dysbiosis include inflammatory bowel disease, obesity, insulin resistance, type II diabetes, and allergies. Kojibiose also has a low glycemic index and is considered an anti-diabetic sweetener (Non-Patent Document 6).

WO2017/218680 (Special Publication No. 2019-517828) WO2018/220237 (Special Publication No. 2020-521801) WO2019/156234 (Special Publication No. 2021-512638) JP 2021-138738 A WO2019/191390 (Special Publication No. 2021-519299) JP 2021-195334 A JP 2005-281188 A (Patent No. 4746281 A) WO2016/172658 (Special Publication No. 2018-513196)

Nat Med. 2019;25(7):1164-1174. J Appl Microbiol. 2021;131(6):3018-3031. Diabetes Ther. 2017;8(2):293-307. PLoS One. 2016;11(2):e0149564. Journal of agricultural and food chemistry. 2005, 53, 5192-5199. Bioprocess Biosyst Eng. 2018 Aug;41(8):1121-1131.

Since there are many reports that intestinal bacteria are closely related to health, it may be possible to reduce the risk of various diseases by controlling the growth of important bacteria.

By controlling the proliferation of Subdoligranulum bacteria in the intestines with prebiotics, it is expected that various diseases can be prevented or improved.

The present invention provides the following:
[1] A composition for controlling the growth of bacteria belonging to the genus Subdoligranulum, comprising any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar.
[2] The composition described in 1 for use as a prebiotic or synbiotic.
[3] A composition for treating any condition selected from food allergies, inflammatory bowel disease, obesity, dyslipidemia, insulin resistance, and diabetes, comprising kojibiose and an oligosaccharide having kojibiose as a constituent saccharide.
[4] The composition described in 3, wherein the treatment is mediated by controlling the proliferation of Subdoligranulum bacteria in the intestine.
[5] A composition for treating a disease or condition that is ameliorated by controlling the proliferation of intestinal bacteria of the genus Subdoligranulum, comprising kojibiose and oligosaccharides having kojibiose as a constituent sugar.
[6] The composition according to any one of 1 to 5, which is obtained by allowing glucansucrase to act on a raw material composition containing lactose and sucrose.
[7] The composition according to any one of claims 1, 2, and 4 to 6, wherein the growth control is growth promotion.
[8] The composition according to any one of 1 to 7, comprising galactosyl kojibiose as a constituent saccharide selected from kojibiose and oligosaccharides having kojibiose.
[9] Obtaining information about a subject's intestinal microbiota;
Deriving information on foods to be provided to a subject based on the information on the intestinal microbiota; and providing the derived information on foods to a subject,
A food information providing method in which, in the information acquiring step, information on the presence or absence or amount of Subdoligranulum bacteria is acquired from information on the intestinal bacterial flora, and, in the food information deriving step, information on a food that is a composition containing any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent sugar is derived based on the information on the presence or absence or amount of Subdoligranulum bacteria.
[10] Carrying out the method according to claim 10,
A method for controlling Subdoligranulum bacteria in the intestines of a subject, or a method for supporting the diet or health of a subject, comprising a step of providing food to the subject based on the derived food information.
[11] The method according to claim 9 or 10, wherein the food is for use as a prebiotic or synbiotic.
[12] The method according to any one of claims 9 to 11, further comprising a step of displaying the provided food information on a target terminal.
[13]
2. Use of any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar, for controlling the proliferation of bacteria of the genus Subdoligranulum in intestinal flora.

[21] A composition comprising any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar, for use in a method for controlling the growth of bacteria of the genus Subdoligranulum. Use of any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent sugar, in the manufacture of a composition for controlling the growth of bacteria of the genus Subdoligranulum. A method or non-therapeutic method for controlling the growth of bacteria of the genus Subdoligranulum, comprising a step of administering to a subject a composition comprising any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent sugar. Use or non-therapeutic use of a composition comprising any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent sugar, for controlling the growth of bacteria of the genus Subdoligranulum.
[22] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to 21, wherein the composition is for use as a prebiotic or synbiotic.
[23] A composition comprising any one selected from kojibiose and oligosaccharides having kojibiose as a constituent saccharide for use in a method for treating any one selected from obesity, dyslipidemia, insulin resistance, diabetes, food allergies, and inflammatory bowel disease. Use of any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide in the manufacture of a composition for treating any one selected from obesity, dyslipidemia, insulin resistance, diabetes, food allergies, and inflammatory bowel disease. A method or non-therapeutic method for treating any one selected from obesity, dyslipidemia, insulin resistance, diabetes, food allergies, and inflammatory bowel disease, comprising the step of administering to a subject a composition comprising any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide. 2. Use or non-therapeutic use of a composition comprising any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent sugar, for treating any one selected from obesity, dyslipidemia, insulin resistance, diabetes, food allergy, and inflammatory bowel disease.
[24] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to 23, wherein the treatment is mediated by controlling the growth of bacteria of the genus Subdoligranulum.
[25] A composition comprising any one selected from kojibiose and oligosaccharides having kojibiose as a constituent saccharide, for use in a method for treating a disease or condition that can be improved by controlling the growth of intestinal bacteria of the genus Subdoligranulum. Use of any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide, in the manufacture of a composition for treating a disease or condition that can be improved by controlling the growth of intestinal bacteria of the genus Subdoligranulum. A method or non-therapeutic method for treating a disease or condition that can be improved by controlling the growth of intestinal bacteria of the genus Subdoligranulum, comprising a step of administering to a subject a composition comprising any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide. Use or non-therapeutic use of a composition comprising any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide, for treating a disease or condition that can be improved by controlling the growth of intestinal bacteria of the genus Subdoligranulum.
[26] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of claims 21 to 25, wherein the composition is obtained by acting on a raw material composition containing lactose and sucrose with glucansucrase.
[27] The composition, the use in manufacture, the method or non-therapeutic method, or the use or non-therapeutic use according to any one of claims 21, 22, 24 to 26, wherein the growth control is growth promotion.
[28] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 21 to 27, wherein the composition contains galactosyl kojibiose as any one selected from kojibiose and oligosaccharides having kojibiose as a constituent saccharide.
[29] A method for controlling the proliferation of bacteria of the genus Subdoligranulum in an intestinal bacterial flora, the method comprising a step of causing an intestinal bacterial flora containing bacteria of the genus Subdoligranulum to contain any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar.

By using kojibiose or an oligosaccharide containing kojibiose as a prebiotic, it is possible to increase the number of Subdoligranulum bacteria in the intestines. Furthermore, by increasing the number of Subdoligranulum bacteria in the intestines, it is expected that various diseases can be prevented and improved.

Changes in the occupancy rate of Subdoligranulum bacteria in the V3-V4 region amplicon sequence by MiSeq (Illumina) using galactosyl kojibiose (trisaccharide fraction) in a human fecal culture system. Black circles indicate the progress of each subject, and horizontal bars indicate the average value. Chromatograms of the enzyme reaction mixture and the trisaccharide and tetrasaccharide fractions after fractionation and purification

This embodiment relates to a composition for controlling the growth of bacteria of the genus Subdoligranulum, represented by Subdoligranulum, which contains kojibiose and oligosaccharides having kojibiose as its constituent sugars as active ingredients.

[Active ingredient]
The composition of the present embodiment contains, as an active ingredient, any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide. When referring to oligosaccharides having kojibiose as a constituent saccharide, kojibiose itself is not included. In addition, in the present invention, "any one" is used to mean "at least one" unless otherwise specified. For example, "any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent saccharide" includes only kojibiose, only one type of oligosaccharide having kojibiose as a constituent saccharide, one type of oligosaccharide having kojibiose and kojibiose as a constituent saccharide, two types of oligosaccharides having kojibiose as a constituent saccharide, etc. "Containing as an active ingredient" refers to the fact that the active ingredient is used in the composition in an effective amount to exert the intended function, that the active ingredient is specified by labeling as an ingredient that contributes to the intended purpose, and the like. In foods with functional claims, active ingredients are sometimes referred to as functional ingredients (ingredients that contribute to specific health purposes (excluding those related to reducing the risk of disease)).

The kojibiose and oligosaccharides having kojibiose as a constituent sugar used in the composition are not particularly limited as long as they have the intended effect. In addition, the kojibiose and oligosaccharides having kojibiose as a constituent sugar used in the composition may be one type or a combination of two or more types.

Examples of oligosaccharides that contain kojibiose include the trisaccharides galactosyl kojibiose, kojitriose, selaginose (kojibiosylglucoside), centose, and kojibiosylfructoside, as well as tetrasaccharides in which one molecule of a monosaccharide such as glucose is bonded to each of these trisaccharides, such as kojitetraose and kojitriosylglucoside.

In a preferred embodiment, the composition contains either kojibiose or galactosyl kojibiose. Examples of such compositions include a composition containing kojibiose, a composition containing galactosyl kojibiose, a composition containing kojibiose and galactosyl kojibiose, and a composition containing kojibiose, galactosyl kojibiose, and a tetrasaccharide in which one glucose molecule is bound to galactosyl kojibiose.

In the context of this invention, unless otherwise specified, galactosyl kojibiose refers to kojibiose in which galactose is α- or β-linked to any of positions 1 to 8 of the following kojibiose. Theoretically, galactose can be linked to the OH at position 8 in four ways (α-α, α-β, β-α, β-β) and to the OH at positions 2 to 7 in two ways, so 18 types of galactosyl kojibiose are expected. They are called 8α-α kojibiose, 8α-β kojibiose, 8β-α kojibiose, 8β-β kojibiose, 1α kojibiose, 2β kojibiose, 2α kojibiose, 2β kojibiose, 3α kojibiose, 3β kojibiose, 4α kojibiose, 4β kojibiose, 5α kojibiose, 5β kojibiose, 6α kojibiose, 6β kojibiose, 7α kojibiose, and 7β kojibiose, respectively.

In one embodiment, the composition includes any one of the 18 types of galactosyl kojibiose, namely, 8α-α kojibiose, 8α-β kojibiose, 8β-α kojibiose, 8β-β kojibiose, 1α kojibiose, 2β kojibiose, 2α kojibiose, 2β kojibiose, 3α kojibiose, 3β kojibiose, 4α kojibiose, 4β kojibiose, 5α kojibiose, 5β kojibiose, 6α kojibiose, 6β kojibiose, 7α kojibiose, and 7β kojibiose, and may include 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18.

In a preferred embodiment, the galactosyl kojibiose contained in the composition is α-D-glucopyranosyl-(1→2)-[β-D-galactopyranosyl-(1→4)-]D-glucopyranoside. This sugar can also be expressed as a type of glucosyl lactose.

According to the studies of the present inventors, galactosyl kojibiose has been confirmed to have an excellent effect of controlling the proliferation of Subdoligranulum bacteria, which is a type of bacteria of the genus Subdoligranulum. Here, galactosyl kojibiose is a type of oligosaccharide that contains kojibiose as a constituent sugar, and after being administered to a subject, it can be broken down by lactase in the subject's digestive tract to produce kojibiose. Therefore, any sugar that contains kojibiose as a constituent sugar can similarly produce kojibiose in the intestines and exert the same effect as galactosyl kojibiose.

The kojibiose and oligosaccharides having kojibiose as a constituent sugar used in the present invention are not particularly limited in terms of origin or production method, and may be produced by any method, such as fermentation, enzymatic or organic synthesis. One preferred production method includes a production method including a step of reacting a raw material composition containing lactose and sucrose with glucansucrase, or a lactic acid bacterium expressing glucansucrase, or a processed product thereof, to obtain a composition containing galactosyl kojibiose. In the present invention, glucansucrase refers to an enzyme belonging to glycoside hydrolase (also called glycosyl hydrolases) family 70 (GH70), unless otherwise specified (see (CAZy) databases and Cantarel et al., Nucleic Acids Res. 37:D233-238, 2009).

When lactic acid bacteria are used, they are preferably lactic acid bacteria that express glucansucrase.Preferred examples of lactic acid bacteria include lactic acid bacteria belonging to the family Leuconostoc, more preferably lactic acid bacteria belonging to the genus Leuconostoc, and even more preferably Leuconostoc mesenteroides.
When using lactic acid bacteria, it is preferable that the lactic acid bacteria constantly express glucansucrase.Preferred examples of lactic acid bacteria are lactic acid bacteria belonging to the Lactobacillaceae family, more preferably lactic acid bacteria belonging to the Liquorilactobacillus genus or lactic acid bacteria belonging to the Limosilactobacillus genus, and even more preferably Limosilactobacillus reuteri and Liquorilactobacillus satsumensis.
When using lactic acid bacteria, it is preferable that the lactic acid bacteria constantly express glucansucrase, and more preferably, the glucansucrase is of the cell-bound type. A preferred example of lactic acid bacteria is a lactic acid bacteria belonging to the Lactobacillaceae family, more preferably, a lactic acid bacteria belonging to the Liquorilactobacillus genus, and even more preferably, Liquorilactobacillus satsumensis. A particularly preferred example of Liquorilactobacillus satsumensis strain is JCM12392. Note that JCM12392 is written as Lactobacillus satsumensis in RIKEN BioResource Center, GENERAL CATALOG No.9, 2012, JAPAN COLLECTION OF MICROORGANISMS, M51.

In relation to the present invention, when describing lactic acid bacteria, unless otherwise specified, the following is used: Zheng J, Wittouck S, Salvetti E, Franz CMAP, Harris HMB, Mattarelli P, O'Toole PW, Pot B, Vandamme P, Walter J, Watanabe K, Wuyts S, Felis GE, Ganzle MG, Lebeer S.: A taxonomic note on the genus Lactobacillus: Description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. According to the reclassification by Int J Syst Evol Microbiol. 2020 Apr; 70(4): 2782-2858. Liquorilactobacillus satsumensis would be classified as Lactobacillus satsumensis according to the previous classification.

The lactic acid bacteria expressing glucansucrase or a processed product thereof is preferably any one selected from the group consisting of live bacteria, killed bacteria, a culture containing bacteria, a disrupted bacteria product, and a purified product of glucansucrase.

Glucansucrase, or lactic acid bacteria expressing glucansucrase, or processed products thereof, used for producing kojibiose and oligosaccharides having kojibiose as a constituent sugar can be produced using transformation, chemical synthesis, or genome editing techniques.

The composition containing galactosyl kojibiose obtained by such a production method can be used as it is as a solution containing kojibiose and oligosaccharides whose constituent sugars are kojibiose, or after purification with an ion exchange resin or the like. In other words, the composition of the present invention may be a composition containing galactosyl kojibiose obtained by acting a composition containing sucrose and lactose with glucansucrase, or a lactic acid bacterium expressing glucansucrase, or a processed product thereof.

[Application]
(Functions, Actions, Effects)
The composition of the present embodiment can be used to control the growth of bacteria belonging to the genus Subdoligranulum (sometimes simply referred to as Subdoligranulum bacteria). In a preferred embodiment, the composition can be used to control the growth of Subdoligranulum bacteria in the human intestine. Controlling the growth of Subdoligranulum bacteria in the human intestine refers to controlling the growth of Subdoligranulum bacteria in a state in which various bacteria living in the human intestine are present (sometimes referred to as intestinal flora). In the present invention, the term intestinal flora refers to a flora in which various bacteria living in the intestine are present, including, for example, a flora contained in feces, and is not limited to a flora in the intestine. In the present invention, control includes upward control (sometimes referred to as promotion or increase) and downward control (sometimes referred to as suppression or decrease), and is preferably upward control. The control of growth can be the control of the number of bacteria or the control of the occupancy rate described later.

In the context of the present invention, the intestine refers to the digestive organ in humans and animals where bacteria reside and which digests and absorbs ingested food. The intestine includes the small intestine and large intestine, and is preferably the large intestine.

In a preferred embodiment, the composition is used to control the proliferation of bacteria belonging to the genus Subdoligranulum (sometimes simply referred to as Subdoligranulum bacteria) in the intestine.

In the context of the present invention, the term "Subdoligranulum bacteria" refers to bacteria that are identified as belonging to the genus Subdoligranulum by molecular phylogenetic analysis based on the 16S rRNA gene. The criteria for determining genera by molecular phylogenetic analysis based on the 16S rRNA gene are well known to those skilled in the art (Stackebrandt E, Ebers J. Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 2006;33:152-155.).

In the context of the present invention, controlling the proliferation of a specific bacterium in a bacterial flora means controlling the proportion (occupancy rate) of the specific bacterium in the bacterial flora.

Whether or not a certain component controls the proliferation of Subdoligranulum bacteria in the intestine can be evaluated as follows: Feces containing Subdoligranulum bacteria provided by a healthy individual is added to an appropriate culture medium and, if necessary, cultivated for a certain period of time, after which the component to be evaluated is added and cultivated under appropriate conditions (e.g., cultivation for 48 hours at 37°C under anaerobic conditions, which are similar to the conditions in the intestine). The number of Subdoligranulum bacteria in the bacterial flora in the culture is then measured. The measurement results can then be compared with those of a culture cultivated under identical conditions, except that a control (e.g., sterilized water) was added instead of the component to be evaluated, to make a judgment.

The number of bacteria of the genus Subdoligranulum contained in the bacterial flora can be determined by a known method. One of the preferred methods is to perform 16S metagenomic analysis (sequence analysis of 16S rRNA gene amplicon) on DNA extracted from the culture. When performing 16S metagenomic analysis, DNA can be extracted using a commercially available kit. The genomic region to be analyzed is not particularly limited as long as it can identify bacteria, but the V3-V4 region of the 16S rRNA gene can be used. Methods such as primers, amplification conditions, and amplicon purification for analyzing bacteria can also be methods well known to those skilled in the art. It is preferable to decipher the sequence using a next-generation sequencer with higher performance. A next-generation microbiome bioinformatics platform such as QIIME 2 ^TM can be used to analyze the obtained data. For 16S metagenomic analysis, those skilled in the art can refer to information such as Sanschagrin S, Yergeau E. Next-generation sequencing of 16S ribosomal RNA gene amplicons. J Vis Exp. 2014;(90):51709. Published 2014 Aug 29. doi:10.3791/51709.

In one aspect, the composition can be used to treat any of the following conditions: food allergy, inflammatory bowel disease, obesity, dyslipidemia, insulin resistance, and diabetes. The treatment is not limited to treatment via regulation of the proliferation of Subdoligranulum bacteria in the intestine, unless otherwise specified.

Subdoligranulum bacteria are bacteria that live in the intestines of mammals, including humans. They are known to be less prevalent in children with food allergies and in patients with inflammatory bowel disease (Non-Patent Documents 1 and 2, supra). It has also been shown that administering Subdoligranulum bacteria to food allergy model mice can suppress food allergy symptoms (Non-Patent Document 1, supra), and it is known that Subdoligranulum bacteria show a positive correlation with HDL cholesterol (Non-Patent Document 3, supra) and a negative correlation with insulin resistance, body weight, and C-reactive protein (CRP), whose blood concentration rapidly increases when inflammation or tissue destruction is present (Non-Patent Document 4, supra). Therefore, in one embodiment, the composition can be used for any treatment selected from the group consisting of obesity, lipid metabolism disorders, insulin resistance, diabetes, food allergies, and inflammatory bowel disease, which is a treatment mediated by controlling the proliferation of Subdoligranulum bacteria in the intestine.

Obesity is a condition in which excess fat accumulates in the body (BMI of 25 or higher). When obesity leads to diseases that have a negative impact on health, or when obesity leads to visceral obesity, it is called obesity. Glucose metabolism disorder refers to a state in which glucose metabolism is abnormal. Lipid metabolism disorder, also known as dyslipidemia or hyperlipidemia, refers to a state in which there is too much LDL cholesterol or triglyceride in the blood, or a state in which HDL cholesterol is low. Lipid metabolism disorders include hyper-LDL cholesterolemia (140 mg/dL or higher), borderline hyper-LDL cholesterolemia (120-139 mg/dL), hypertriglyceridemia (150 mg/dL or higher), and hypo-HDL cholesterolemia (less than 40 mg/dL). Insulin resistance refers to a state in which, despite insulin being secreted into the blood from the pancreas, the sensitivity of target organs to insulin decreases, and its action becomes sluggish. In a state of insulin resistance, the glucose uptake ability of muscle and fat tissues is reduced, and gluconeogenesis cannot be suppressed in the liver, so blood glucose levels are difficult to lower, and more insulin is required to return blood glucose levels to normal. If this condition continues, the insulin secretion function of the pancreas decreases, and blood glucose levels rise, which is said to cause type II diabetes. Insulin resistance can be caused by genetics, obesity, lack of exercise, a high-fat diet, or stress. Diabetes is a disease in which hyperglycemia continues chronically due to insufficient insulin action, and includes type 1 diabetes and type 2 diabetes. Food allergies are caused by ingested foods and cause symptoms such as hives, eczema, diarrhea, and coughing through immunological mechanisms. Food allergies include systemic allergies as well as oral allergy syndrome, which often only has symptoms in the mouth. Inflammatory bowel diseases include ulcerative colitis and Crohn's disease.

When used for the treatment of inflammatory bowel disease, the composition can be configured to be free of any of the following:
Oligosaccharides (e.g., raffinose, stachyose, galactooligosaccharides, isomaltooligosaccharides, lactofructose oligosaccharides, lactulose, xylooligosaccharides, agarooligosaccharides, mannooligosaccharides, or fructooligosaccharides), inulin, pectin, processed pectin, guar gum, decomposed guar gum, psyllium seed gum, karaya gum, tragacanth gum, gum arabic, resistant starch, resistant dextrin, polydextrose, cellulose, hemicellulose, soybean polysaccharides, β-glucan, glucomannan, galactomannan, chondroitin sulfate, hyaluronic acid, levan, lignin, alginic acid and its salts, agarose, and chitosan.

The composition can also be used to treat diseases or conditions that can be improved by controlling the proliferation of Subdoligranulum bacteria in the intestinal tract. Such diseases or conditions can include obesity, dyslipidemia, insulin resistance, diabetes, food allergies, inflammatory bowel disease, and a variety of other conditions.

In the present invention, when referring to a disease or condition, the term "treatment" includes reducing the risk of onset, delaying onset, prevention, treatment, and halting or delaying progression. Treatment also includes maintaining a normal or desirable state, maintaining ability, and alleviating symptoms (preferably temporary alleviation of symptoms). Treatment further includes helping to improve from a disease or condition, alleviating a disease or condition, and reducing the risk of developing a disease or condition. Treatment includes radical treatment (treatment to remove the cause) and symptomatic treatment (treatment to improve symptoms). Actions for improvement or treatment include medical actions performed by doctors and nurses and midwives under the instructions of doctors, as well as non-therapeutic actions performed by persons other than doctors, such as pharmacists, nutritionists (including registered dietitians and sports nutritionists), public health nurses, midwives, nurses, clinical laboratory technicians, sports instructors, pharmaceutical manufacturers, pharmaceutical distributors, food manufacturers, food distributors, etc. Prevention or reduction of the risk of onset further includes recommendations for the intake of specific foods and nutritional guidance (including nutritional guidance necessary for the medical treatment of injured or sick people, and nutritional guidance for maintaining and promoting health).

(subject)
The composition of the present embodiment is intended for subjects in whom it is desirable or necessary to control the proliferation of Subdoligranulum bacteria in the intestine; subjects at risk of any of obesity, dyslipidemia, insulin resistance, diabetes, food allergies, and inflammatory bowel disease; and subjects in whom it is desirable or necessary to treat any of obesity, dyslipidemia, insulin resistance, diabetes, food allergies, and inflammatory bowel disease.

Subjects for whom it is desirable or necessary to control the proliferation of Subdoligranulum bacteria in the intestines include subjects for whom relatively low levels of Subdoligranulum bacteria have been detected in at least one of the intestines and feces. Subjects for whom it is desirable or necessary to control the proliferation of Subdoligranulum bacteria in the intestines include subjects for whom the number or occupancy rate of Subdoligranulum bacteria in the intestines is relatively low through any test or analysis, such as an intestinal flora test, a test for substances in the feces, or a fecal metabolome analysis, subjects for whom it is deemed that there is a high risk of any of the following conditions selected from obesity, dyslipidemia, insulin resistance, diabetes, food allergies, and inflammatory bowel disease, and subjects for whom the intake of food, drink, or medicine that controls the proliferation of Subdoligranulum bacteria in the intestines has been suggested by a doctor, nurse, pharmacist, nutritionist, etc., or a computer program based on the results of the test or analysis.

　Subjects in which a relatively low number or occupancy rate of Subdoligranulum bacteria has been detected in at least one of the intestines and feces include, for example, cases in which, when a culture solution containing stool collected from a subject is cultured at 37°C under anaerobic conditions for 24 hours, the occupancy rate of Subdoligranulum bacteria in the culture solution is 2% or less, 1.75% or less, 1.5% or less, 1.25% or less, 1% or less, 0.75% or less, 0.5% or less, 0.4% or less, 0.3% or less, or 0.3% or less.

In one embodiment, the subject is healthy (not diagnosed with a disease by a physician). The subject may be a human or a non-human animal.

The age of the subject is not particularly limited, and the subject may be, for example, a newborn (within 28 days of birth); an infant (less than 1 year of age); a toddler (1-6 years of age); a child (7 years of age or older, but under 15 years of age); an adult (15 years of age or older); or a person aged 65 years or older.

[Composition]
(Food composition, etc.)
The composition of the present invention can be a food composition or a pharmaceutical composition. Food and pharmaceutical products include not only those for humans but also those for animals other than humans, unless otherwise specified. Food includes general food, functional food, nutritional composition, and also therapeutic food (for the purpose of treatment. Prepared based on a menu prepared by a nutritionist or the like according to a doctor's meal prescription), dietary therapy food, ingredient-adjusted food, nursing care food, and food for medical support, unless otherwise specified. Food includes not only solid food but also liquid food, such as beverages, energy drinks, liquid food, and soup, unless otherwise specified. Functional foods refer to foods that can impart a certain functionality to the living body, and include, for example, foods for specified health uses (including conditionally designated health foods [specified health foods]), functional foods, health functional foods including nutritional functional foods, special purpose foods, nutritional supplements, dietary supplements, supplements (for example, tablets, coated tablets, sugar-coated tablets, enteric preparations, capsules (enteric soft capsules, enteric hard capsules, colon delivery capsules, etc.), liquids, and other dosage forms), dietary supplements, food supplements, medical foods (following the definition of the Food and Drug Administration (FDA)), beauty foods (for example, diet foods), and other health foods in general. In addition, in the present invention, "functional foods" includes health foods to which a health claim based on the food standards of Codex (FAO/WHO Joint Food Standards Commission) is applied. Food supplements are supplements to normal meals and are concentrated with nutrients or other substances that have nutritional or physiological effects, either alone or in combination, and are labeled as Food Supplements. A dietary supplement is a product (excluding tobacco) intended to supplement the diet, which contains one or more of the targeted ingredients and is labeled as a dietary supplement. The composition may also be a refined product of a specific sugar. Refined products include fractions, partially refined products, and crude refined products. When actually labeling food products, the labeling laws of each country must be followed.

(Route of administration, etc.)
The composition of the present invention may be administered orally, parenterally, for example, through a tube (gastrostomy, enterostomy), or nasally, but is preferably administered orally. In the present invention, administration is used not only when a pharmaceutical product is administered to a subject, but also when a food other than a pharmaceutical product is ingested by a subject. "Administer" can be read as "intake," and "administer" can be read as "administer."

The composition can be administered to a subject repeatedly, and can be administered to a subject continuously for an extended period of time. The period is not particularly limited, but in order to fully demonstrate the effect, it is preferable to administer the composition continuously for a relatively long period of time, for example, 3 days or more, 1 week or more, 2 weeks or more, 1 month or more, 3 months or more, 6 months or more, or 1 year or more.

The composition may be administered routinely, proactively, such as when there is a high risk, or when the need arises. The composition may be administered with a meal, before, after, or between meals, or at the onset of the disease or condition that it is desired to ameliorate with the composition.

(dose, content)
In this embodiment, the active ingredient kojibiose or any one selected from oligosaccharides having kojibiose as a constituent sugar can be used in an amount acceptable for food or medicine. The dosage of the composition of this embodiment may be any amount that exerts the desired effect. The dosage can be appropriately set in consideration of various factors such as the age, weight, and symptoms of the subject.

The daily dosage of the composition may be 0.5 g or more, 1 g or more, preferably 2 g or more, preferably 3 g or more, more preferably 5 g or more, and even more preferably 10 g or more, in terms of the amount of active ingredient. The upper limit of the amount of active ingredient per day, whatever the lower limit, may be 80 g or less, 70 g or less, 60 g or less, 50 g or less, 40 g or less, 30 g or less, 20 g or less, or 15 g or less. When the composition contains multiple active ingredients, the amount of active ingredient refers to the total amount of active ingredients contained.

Dosage may be once a day or multiple times a day, for example 2 to 10 times. The amount of active ingredient per dose may be, for example, 0.5 g or more, 1 g or more, preferably 2 g or more, more preferably 3 g or more, and even more preferably 5 g or more. The upper limit of the amount of active ingredient per dose, whatever the lower limit, may be 70 g or less, 60 g or less, 50 g or less, 40 g or less, 30 g or less, 25 g or less, or 10 g or less.

The content of the active ingredient in the composition may be an amount acceptable for use as a food or medicine, and may be set appropriately depending on the form of the composition. For example, when the composition is in the form of a food or medicine to be eaten or consumed as is, such as fermented milk or a beverage, the content of the active ingredient per 100 g of the composition may be 0.01% or more, preferably 0.1% or more, more preferably 0.3% or more, and even more preferably 0.5% or more. The upper limit of the active ingredient per 100 g of the composition, whatever the lower limit, may be 8% or less, 5% or less, 4% or less, or even 3% or less. Alternatively, the content of the active ingredient per solid content of the composition may be 0.1% or more, preferably 1% or more, more preferably 3% or more, and even more preferably 5% or more. The upper limit of the active ingredient per solid content, whatever the lower limit, may be 80% or less, 50% or less, 40% or less, or 30% or less. In the present invention, % means % by mass, unless otherwise specified.

(Other ingredients, additives)
In the present invention, the composition may contain other active ingredients or nutritional ingredients that are acceptable as foods or pharmaceuticals. Examples of such ingredients include lipids (e.g., milk fat, vegetable oils and fats, oils and fats containing medium-chain fatty acids), proteins (e.g., milk proteins, milk protein concentrates (MPC), whey protein concentrates (WPC), whey protein isolates (WPI), α-lactalbumin (α-La), β-lactoglobulin (β-Lg), heat-denatured whey proteins, and enzyme-treated whey proteins), amino acids (e.g., lysine, arginine, glycine, alanine, glutamic acid, leucine, isoleucine, valine), kojibiose, and sugars containing kojibiose as a constituent sugar. Examples of suitable nutrients include carbohydrates other than oligosaccharides (glucose, sucrose, fructose, maltose, trehalose, erythritol, maltitol, palatinose, xylitol, dextrin), electrolytes (e.g., sodium, potassium, calcium, magnesium), vitamins (e.g., vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid, and nicotinic acids), minerals (e.g., copper, zinc, iron, cobalt, manganese), antibiotics, and dietary fiber.

　In the present invention, the composition may contain a prebiotic other than the active ingredient. Prebiotics can be defined as indigestible food ingredients that have a beneficial effect on the host and improve the host's health by selectively altering the growth and activity of specific bacteria in the large intestine. The composition may contain one type or two or more types of prebiotics other than the active ingredient.

　There are no particular limitations on prebiotics other than the active ingredient, so long as they do not interfere with the effects of the active ingredient contained in the composition. Examples of prebiotics other than the active ingredient include galactooligosaccharides, fructooligosaccharides (GF2, GF3, GF4), xylooligosaccharides, isomaltooligosaccharides, raffinose, lactulose, lactosucrose, soybean oligosaccharides, coffee oligosaccharides, dietary fiber, and gluconic acid.

The composition can also be used as a synbiotic, and may contain probiotics in addition to the active ingredient. Synbiotics are a combination of probiotics and prebiotics. Probiotics can be defined as microorganisms that have a beneficial effect on the host when introduced into the intestine of the host in a live state.

There are no particular limitations on the probiotics, so long as they do not interfere with the effects of the active ingredients contained in the composition. Certain lactic acid bacteria are known as examples of probiotics.

The composition may further contain additives that are acceptable for use as food or medicine. Examples of such additives include inert carriers (solid or liquid carriers), excipients, surfactants, binders, disintegrants, lubricants, solubilizers, suspending agents, coating agents, colorants, preservatives, buffers, pH adjusters, emulsifiers, stabilizers, sweeteners, antioxidants, flavors, acidulants, and natural products. More specifically, these include water, other aqueous solvents, pharma- ceutical acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium alginate, dextrin, water-soluble dextran, water-soluble dextrin, corn starch, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, magnesium stearate, precipitated calcium carbonate, talc, human serum albumin, mannitol, sorbitol, lactose, sucralose, lactose hydrate, candy powder, white sugar, stevia, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, vegetable juice, etc.

(Dosage form/shape)
In one embodiment, the food composition may be prepared in any form, such as a solid, liquid, mixture, suspension, powder, granule, paste, jelly, gel, capsule, etc. The food composition according to the present invention may be in any form, such as a dairy product, a supplement (e.g., a tablet, a coated tablet, a sugar-coated tablet, an enteric coating, etc., a capsule (enteric soft capsule, enteric hard capsule, large intestine delivery capsule, etc.), a confectionery, a beverage, a drink, a seasoning, a processed food, a side dish, a soup, etc. More specifically, the composition according to the present invention may be in the form of a liquid food, a semi-liquid food, a jelly, a gel, a powder, a formula, a liquid formula, a powder or liquid milk for pregnant women and lactating women, a fermented milk, a bar, a mousse, a chocolate, a biscuit, an ice cream, a fermented milk, a lactic acid bacteria drink, a dairy drink, a milk drink, a soft drink, a fruit juice drink, a tablet, a cheese, a bread, a biscuit, a cracker, a pizza crust, a food for the sick, a nutritional food, a frozen food, a processed food, etc. The formula may be in the form of granules, powder, paste, thickened liquid, etc., for mixing with beverages or foods. The granules and powder may be in the form of cubes or sticks (each packaged in a single serving). In the present invention, modified powdered milk refers to raw milk, cow's milk, special cow's milk, raw buffalo milk, or foods made from these ingredients, processed or used as the main ingredient, to which nutrients necessary for infants are added, and made into a powder, as defined in the "Ministerial Ordinance on the Compositional Standards, etc. of Milk and Dairy Products (hereinafter referred to as the "Ministerial Ordinance on Milk, etc."). In the present invention, modified liquid milk refers to raw milk, cow's milk, special cow's milk, raw buffalo milk, or foods made from these ingredients, processed or used as the main ingredient, to which nutrients necessary for infants are added, and made into a liquid, as defined in the Ministerial Ordinance on Milk, etc.

In one embodiment, the pharmaceutical composition can be in any dosage form suitable for oral administration, such as solid preparations such as tablets, granules, powders, pills, and capsules, liquid preparations such as solutions, suspensions, and syrups, gels, and aerosols.

(Manufacturing method, etc.)
In one embodiment, in the production of the composition, the stage of blending the active ingredient can be appropriately selected. The stage of blending is not particularly limited as long as it does not significantly impair the properties of the active ingredient. For example, the active ingredient can be blended by mixing it with the raw material. Alternatively, the active ingredient can be added at the final stage of production to produce a composition containing the active ingredient.

(display)
In one embodiment, the composition can be labeled with its intended use (application), and in another embodiment, the function of the composition or active ingredient or the usage based on the function is displayed. Examples of usage based on the function are as described above for the functions, actions, and effects. In addition, the composition of the present invention can be labeled with the fact that it can be used as a prebiotic or as a synbiotic (a combination of probiotics and prebiotics). Note that a period such as "temporary" or "long-term" may be displayed at the beginning of each wording as appropriate.

In one embodiment, the composition is labeled to recommend administration to a specific subject. Examples of subjects to which the labeling is provided are as described above for subjects.

　Display can be explicit or implicit. An example of explicit display is a direct statement on a tangible object such as the product itself, packaging, container, label, tag, etc., while an example of implicit display (which can also be called implied) includes advertising and promotional activities by place or means such as websites, storefronts, pamphlets, exhibitions, seminars such as media seminars, books, newspapers, magazines, television, radio, mail, email, and voice.

In one embodiment, the recommendation to consume the composition is displayed personally. Such a display can be made using a document (whether written or electronic) addressed to the subject, the subject's tablet, smartphone, personal computer, the subject's SNS, etc. Furthermore, such a display can be made together with the display of the results of any test or analysis, such as an intestinal flora test, a test of substances in the feces, and a fecal metabolome analysis, for the subject. It can also be used for precision nutrition (individualized nutrition, which refers to proposing appropriate meals (nutrition) according to an individual's constitution).

[Providing device, providing method, etc.]
In this embodiment, a food information providing device is provided, comprising:
an information acquisition unit for acquiring information on the intestinal microbiota of a subject;
A derivation unit that derives information on food to be provided to the subject based on the information on the intestinal flora; and a provision unit that provides the derived food information to the subject.

In a preferred embodiment, the information acquisition unit of the device acquires information on the presence or absence or amount of Subdoligranulum bacteria from the intestinal flora information, and the derivation unit derives information on a food that is a composition containing either kojibiose or an oligosaccharide having kojibiose as a constituent sugar, based on the information on the presence or absence or amount of Subdoligranulum bacteria.

In one embodiment, along with obtaining information about the subject's gut microbiota, any of the following information about the subject may be obtained:
- Attribute information of the subject, including any one selected from the group consisting of gender and age;
- Test result information on the subject's physical and health condition, including any selected from the group consisting of height, weight, body mass index (BMI), obesity level, body fat, waist circumference, blood pressure, lipids, liver and pancreas function, metabolic system, blood, urine, renal function, and large intestine;
Survey results information on the subject's preferences and lifestyle, including any of the following: eating habits (preferences), drinking habits, smoking habits, smoking history, exercise habits, and sleep time; and the subject's subjective symptoms, stress, illnesses currently being treated or monitored, medicines and health foods being taken, medical history, pregnancy and childbirth experience; and the subject's work history and general lifestyle, etc.

The information may be obtained based on responses obtained from the subject to predetermined questions (e.g., responses to a questionnaire).

In one aspect, the food information providing device is capable of displaying information about the food to be provided on a target terminal. In another aspect, the device further includes a display unit that displays information about the food to be provided. The display unit may be the target terminal, for example, a tablet terminal, a smartphone, or a personal computer.

In one embodiment, the food information providing device may include an analysis unit that analyzes the bacterial flora of a sample obtained from the subject, and may also include an order receiving unit that receives an order for food from the subject based on the provided food information.

This embodiment also provides a food information providing method, comprising the steps of:
Obtaining information about the gut microbiota of a subject;
Based on the information on the intestinal microbiota, information on foods to be provided to the subject is derived; and the derived food information is provided to the subject.

In a preferred embodiment, in the step of acquiring information, information on the presence or absence or amount of Subdoligranulum bacteria is acquired from information on the intestinal flora, and in the step of deriving food information, information on the food, which is a composition containing any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar, is derived based on the information on the presence or absence or amount of Subdoligranulum bacteria.

Such a method may further include a step of displaying information about the food to be provided on a target terminal. The target terminal may be, for example, a tablet terminal, a smartphone, or a personal computer.

This embodiment also provides a method for controlling Subdoligranulum bacteria in the intestine of a subject or a method for supporting the diet or health of a subject, comprising carrying out the above-mentioned food information providing method and further carrying out the following steps:
A method for controlling Subdoligranulum bacteria in the intestines of a subject, or a method for supporting the diet or health of a subject, comprising a step of providing food to the subject based on the derived food information.

The food in this embodiment is preferably intended for use as a prebiotic or synbiotic. The food here can also be called a food composition, and the explanation in the composition section above applies as is.

[Culture method, etc.]
In another embodiment, there is provided a method for culturing bacteria of the genus Subdoligranulum using any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar, or a use of any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar in culturing bacteria of the genus Subdoligranulum. The culturing or use may be performed in an intestinal flora. In another embodiment, there is provided a method for controlling the proliferation of bacteria of the genus Subdoligranulum in an intestinal flora, the method comprising a step of including any one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar in an intestinal flora containing bacteria of the genus Subdoligranulum. The step of including can be achieved by adding, providing, strengthening, supplementing, or the like, a target component.

The present invention will be explained in more detail below using examples. However, the technical scope of the present invention is not limited to these examples.

[Culture example 1]
(fecal culture)
Feces from healthy adults were suspended in semi-solid medium for GAM glycolysis "Nissui" (Nissui Pharmaceutical Co., Ltd.) from which the agar had been removed by filtration, and galactosyl kojibiose ((α-D-Glcp-(1→2)-[β-D-Galp-(1→4)-]D-Glcp) was added to a concentration of 0.25% (n=60).

Faecal matter from 60 individuals was promptly dispensed under refrigerated conditions after excretion, and then frozen and stored at -80°C until use. These fecal samples were used and tested individually without mixing. The culture medium used in the tests was left to stand at least overnight in an anaerobic glove box before use. A 96-well deepwell plate was used for culture, and the samples were cultured in an anaerobic environment at 37°C for one day. The prebiotic potential of the added evaluation substance was evaluated by comparing the bacterial flora after culture with that in a control condition in which distilled water was added instead of galactosyl kojibiose.

(sample)
Galactosyl kojibiose (α-D-Glcp-(1→2)-[β-D-Galp-(1→4)-]D-Glcp) was prepared from the trisaccharide fraction containing galactosyl kojibiose as the main component prepared in Production Example 1.

(Analysis of gut bacteria occupancy rate using next-generation sequencer)
After incubation, DNA was extracted and purified from the diluted fecal matter using the Maxwell RSC PureFood GMO & Authentication Kit (Promega), and amplicon sequencing of the V3-V4 region was performed using MiSeq (Illumina). The resulting .fastq files were analyzed using QIIME2 (https://qiime2.org/) to calculate the occupancy rate of each enterobacteria.

When galactosyl kojibiose was added, a significant increase in the occupancy rate of Subdoligranulum bacteria was observed (Figure 1, Wilcoxon signed rank test, p = ^3.42x10-4 ).

[Production Example 1]
(Preparation of enzyme solution)
Lactobacillus satsumensis JCM12392 was cultured overnight at 30°C using commercially available MRS liquid medium, and then cooled to below 10°C to terminate the culture. The JCM12392 strain is available from the Japan Collection of Microorganisms, RIKEN BioResource Center (RIKEN BRC) (RIKEN BRC-JCM, Japan) under the JCM number JCM12392.

The resulting culture medium was centrifuged and the supernatant discarded to collect the bacteria, after which an equal volume of citrate phosphate buffer (pH 5.0) was added and the bacteria were collected again to wash the bacteria. The above buffer was added to the obtained bacteria in an amount half that of the original culture medium, and the resulting suspension was used as the enzyme solution.

(Enzyme reaction)
Sucrose (32 w/w%), lactose (16 w/w%), and enzyme solution (10 w/w%) were mixed and reacted at 48°C for 18 hours under static conditions. The composition contained 25-30% galactosyl kojibiose, 15-20% lactose, and 30-35% fructose based on the solid content.

(Purification of Galactosyl Kojibiose)
After passing the enzyme reaction mixture through a carbon celite column, distilled water was passed through to remove non-adsorbed substances. Stepwise elution with aqueous ethanol solutions of varying concentrations yielded a trisaccharide fraction containing galactosyl kojibiose as the main component (Figure 2).

[Summary]
From the above, it can be said that galactosyl kojibiose can be used to control the proliferation of bacteria of the genus Subdoligranulum. In addition, galactosyl kojibiose may generate kojibiose in the intestine by lactase. Therefore, it can be said that kojibiose and oligosaccharides containing kojibiose as a constituent sugar can be used to control the proliferation of bacteria of the genus Subdoligranulum, and further, to treat diseases or conditions that are improved by controlling the proliferation of bacteria of the genus Subdoligranulum.

　Subdoligranulum bacteria are bacteria that live in the intestines of mammals, including humans. It is known that the number of Subdoligranulum bacteria is low in children with food allergies and in patients with inflammatory bowel disease (Non-Patent Documents 1 and 2). It has also been shown that administering Subdoligranulum bacteria to food allergy model mice can suppress food allergy symptoms (Non-Patent Document 1). It is also known that Subdoligranulum bacteria show a positive correlation with HDL cholesterol (Non-Patent Document 3) and a negative correlation with insulin resistance, body weight, and C-reactive protein (CRP), whose blood concentration rapidly increases when there is inflammation or tissue destruction (Non-Patent Document 4). Therefore, it can be said that kojibiose and oligosaccharides containing kojibiose as a constituent sugar can be used to treat these diseases or conditions. It can also be said that the ingestion of a composition containing kojibiose and oligosaccharides containing kojibiose as a constituent sugar is useful for supporting the dietary life of the subject and promoting health.

[Food Manufacturing Example 1: 100% Orange Juice Drink]
168 g of 1/6 concentrated orange juice, 20 g of the oligosaccharide composition (solid content 60%) obtained in Production Example 1, and an appropriate amount of flavoring are dissolved in ion-exchanged water to make a total volume of 1000 ml. This is filled into a container and heat-sterilized at 65°C for 10 minutes to obtain a 100% orange juice beverage containing a trisaccharide fraction containing galactosyl kojibiose as the main component. This juice beverage contains galactosyl kojibiose at about 0.36-0.42 g/100 g. This juice beverage can be used to control the growth of bacteria of the genus Subdoligranulum.

[Food Production Example 2: Drinkable Yogurt]
The raw material cream, skim milk concentrate, and raw material water are mixed and dissolved, heat sterilized at 110°C for 30 seconds, cooled to 43°C, and then a lactic acid bacteria starter consisting of Lactobacillus bulgaricus and Streptococcus thermophilus is added, and the mixture is fermented to an acidity of 0.75. The homogenized solution is mixed and dissolved with raw material water, pectin, sucralose, a trisaccharide fraction containing galactosyl kojibiose as the main component, and flavoring, and heat sterilized at 80°C for 10 minutes. The mixture is mixed and stirred under cooling. The mixing ratio of the raw materials is shown in the table below. The resulting drink yogurt can be used to treat lipid metabolism disorders.

[Food Production Example 3: Fermented Milk 1]
Raw milk is prepared by mixing 500.0 g of raw milk, 53.2 g of skim milk powder, 23.0 g of fresh cream, 403.6 g of tap water, and 50 g of sucrose. The raw milk is heat-sterilized at a temperature of 95°C, and the heat-sterilized raw milk is cooled. Then, the raw milk is inoculated with 0.5% of Liquorilactobacillus satsumensis concentrate, and fermented under static conditions at 48°C for 18 hours, and Lactobacillus bulgaricus and Streptococcus thermophilus are added as lactic acid bacteria starters. The amount of lactic acid bacteria starter added is 20 g. The raw milk to which the lactic acid bacteria starter has been added is filled into a cup container (capacity: 100 ml, made of plastic). The raw milk filled into the cup container is statically fermented in a fermentation room at a temperature of 43°C until the lactic acid acidity reaches 0.7%. The obtained fermented milk can be used for the treatment of lipid metabolism disorders.

[Food Production Example 4: Fermented Milk 2]
Raw milk (500.0 g), skimmed milk powder (53.2 g), fresh cream (23.0 g), tap water (403.6 g), and trisaccharide fraction (54 g) containing galactosyl kojibiose as the main component are mixed to prepare raw milk. The raw milk is then heat-sterilized at 95°C, and the heat-sterilized raw milk is cooled to 43°C. Lactobacillus bulgaricus and Streptococcus thermophilus are then added as lactic acid bacteria starters to the heat-sterilized raw milk. The amount of lactic acid bacteria starter added is 20 g. The raw milk to which the lactic acid bacteria starter has been added is filled into a cup container (capacity: 100 ml, made of plastic). The raw milk filled into the cup container is left to ferment in a fermentation chamber at 43°C until the lactic acid acidity reaches 0.7%. The obtained fermented milk can be used for the treatment of diabetes.

The present invention supports the maintenance and improvement of people's health by using a composition for controlling the proliferation of Subdoligranulum bacteria, which contains any one selected from the group consisting of kojibiose and oligosaccharides whose constituent sugar is kojibiose. The present invention also provides a food composition and a method for producing food that support the maintenance and improvement of people's health. Furthermore, the present invention can improve the nutrition of various people, ensure healthy lives, and promote welfare.

Claims (14)

A composition for controlling the growth of bacteria belonging to the genus Subdoligranulum, comprising either kojibiose or an oligosaccharide having kojibiose as a constituent sugar. The composition of claim 1 for use as a prebiotic or synbiotic. A composition for treating any of food allergies, inflammatory bowel disease, obesity, dyslipidemia, insulin resistance, and diabetes, comprising kojibiose and any of oligosaccharides having kojibiose as a constituent sugar. The composition of claim 3, wherein the treatment is mediated by controlling the proliferation of Subdoligranulum bacteria in the intestine. A composition for treating a disease or condition that is improved by controlling the proliferation of bacteria of the genus Subdoligranulum in the intestine, comprising kojibiose and oligosaccharides having kojibiose as a constituent sugar. The composition according to any one of claims 1 to 5, which is obtained by acting glucansucrase on a raw material composition containing lactose and sucrose. The composition according to any one of claims 1, 2, 4, and 5, wherein the growth control is growth promotion. The composition according to any one of claims 1 to 5, comprising galactosyl kojibiose as one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar. Obtaining information about the gut microbiota of a subject;
Deriving information on foods to be provided to a subject based on the information on the intestinal microbiota; and providing the derived information on foods to a subject,
A food information providing method in which, in the information acquiring step, information on the presence or absence or amount of Subdoligranulum bacteria is acquired from information on the intestinal bacterial flora, and, in the food information deriving step, information on a food that is a composition containing any one selected from the group consisting of kojibiose and oligosaccharides having kojibiose as a constituent sugar is derived based on the information on the presence or absence or amount of Subdoligranulum bacteria. Carrying out the method according to claim 9,
A method for controlling Subdoligranulum bacteria in the intestines of a subject, or a method for supporting the diet or health of a subject, comprising a step of providing food to the subject based on the derived food information. The method according to claim 9 or 10, wherein the food is for use as a prebiotic or synbiotic. The method of claim 9, further comprising a step of displaying the provided food information on a target terminal. 　The use of kojibiose and any of the oligosaccharides having kojibiose as a constituent sugar for controlling the proliferation of Subdoligranulum bacteria in the intestinal flora. A method for controlling the proliferation of bacteria of the genus Subdoligranulum in an intestinal flora, the method comprising a step of causing an intestinal flora containing bacteria of the genus Subdoligranulum to contain one selected from kojibiose and oligosaccharides having kojibiose as a constituent sugar.