Intestinal short-chain fatty acid production promoting composition and food and drink including the same

Abstract

To provide a new functional material which has an effect of promoting production of a short-chain fatty acid in an intestine, in particular, acetic acid, propionic acid, butyric acid, etc.SOLUTION: An intestinal short-chain fatty acid production promoting composition contains: a lactic acid bacterium killed body; and dietary fiber and / or hardly digestible oligosaccharide. The composition is used for increasing an intestinal production amount of one or more short-chain fatty acids selected from the group consisting of acetic acid, propionic acid, and butyric acid. The composition may be used for improving an intestinal bacterial flora. The composition may be a food and drink, a food and drink adding material, a pharmaceutical, a pharmaceutical adding material, animal feed, or an animal feed adding material.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a functional material having an effect of promoting the production of short-chain fatty acids in the intestine, particularly acetic acid, propionic acid, butyric acid and the like.

Background Art

[0002] Food and health are closely related. In recent years, it has been reported that the intestinal bacterial environment is involved in various diseases such as intestinal diseases, immune diseases, metabolic diseases, and mental diseases. Although the intestinal flora and the host maintain homeostasis while acting on each other, the intestinal flora also changes depending on the food ingested. Since intestinal bacteria obtain energy by assimilating oligosaccharides and dietary fibers, which are indigestible components, that reach the large intestine without being digested in the human small intestine, the intestinal environment changes depending on the food ingested by that person.

[0003] In addition, when intestinal bacteria decompose indigestible components, short-chain fatty acids are produced as fermentation products. This short-chain fatty acid not only serves as energy for the large intestinal mucosa, but is also related to sugar metabolism such as improvement of insulin resistance and promotion of insulin secretion-promoting hormone and glucagon peptide-1 (GLP-1) secretion, and improvement of the intestinal environment is effective in preventing lifestyle-related diseases such as diabetes. It has been suggested that there is a possibility.

[0004] On the other hand, focusing on the important role of such short-chain fatty acids, Patent Document 1 discloses an invention of an enteric butyric acid concentration-increasing promoter containing cells of Lactobacillus acidophilus and / or Bifidobacterium longum as an active ingredient.

[0005] In addition, Patent Document 2 discloses an invention of an enteric butyric acid production promoter characterized by containing dead cells of bacteria isolated from the intestine of an animal as an active ingredient.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] However, in Patent Document 1, it is described that when rats are fed a feed containing cells of Lactobacillus acidophilus or Bifidobacterium longum, the butyric acid concentration in the cecum of the rats increases, but short-chain fatty acids other than butyric acid are not clarified.

[0008] Also, in Patent Document 2, it is described that when pigs are fed a diet containing cells of Enterococcus faecalis, the butyric acid concentration in the cecum increases compared to the control, but no significant changes are obtained for acetic acid and propionic acid other than butyric acid, and no change over time in the total short-chain fatty acid concentration is observed (paragraph 0046 of Patent Document 2).

[0009] An object of the present invention is to provide a new functional material having an effect of promoting the production of short-chain fatty acids in the intestine, particularly acetic acid, propionic acid, butyric acid, etc., in view of the prior art.

Means for Solving the Problems

[0010] In order to achieve the above object, as a result of various studies, the present inventors have found that when dead cells of lactic acid bacteria are used in combination with dietary fiber and / or indigestible oligosaccharides, there is an effect of promoting the production of short-chain fatty acids in the intestine, and thus the present invention has been completed.

[0011] That is, the present invention provides a composition for promoting the production of short-chain fatty acids in the intestine, which is characterized by containing dead cells of lactic acid bacteria, dietary fiber and / or indigestible oligosaccharides.

[0012] The dead lactic acid bacteria cells are preferably dead cells of Enterococcus faecalis. Further, the dietary fiber is preferably resistant starch.

[0013] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention can be used to increase the intestinal production amount of one or more short-chain fatty acids selected from the group consisting of acetic acid, propionic acid, and butyric acid.

[0014] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention can be a food or drink, a raw material for adding to food or drink, a pharmaceutical, a raw material for adding to a pharmaceutical, an animal feed, or a raw material for adding to an animal feed.

[0015] From another aspect, the present invention can provide a food or drink containing the above composition for promoting the production of short-chain fatty acids in the intestine.

Effects of the Invention

[0016] According to the present invention, a new functional material having an effect of promoting the production of short-chain fatty acids in the intestine, particularly acetic acid, propionic acid, butyric acid, etc., can be provided by using dead lactic acid bacteria cells and dietary fiber and / or indigestible oligosaccharides.

Brief Description of the Drawings

[0017]

Figure 1

Figure 2

Figure 3

BEST MODE FOR CARRYING OUT THE INVENTION

[0018] The lactic acid bacteria used in the present invention are not particularly limited. For example, microorganisms belonging to the genus Enterococcus such as Enterococcus faecalis, Enterococcus faecium, microorganisms belonging to the genus Lactobacillus such as Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus helveticus, Lactobacillus pentosus, Lactobacillus gasseri, microorganisms belonging to the genus Bifidobacterium such as Bifidobacterium bifidum, microorganisms belonging to the genus Streptococcus such as Streptococcus thermophilus, and microorganisms belonging to the genus Lactococcus such as Lactococcus lactis can be mentioned. Among them, it is preferably one or more selected from the group consisting of Enterococcus faecalis, Enterococcus faecium, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus helveticus, and Lactobacillus pentosus, and particularly preferably Enterococcus faecalis. The lactic acid bacteria may be used alone or in combination of two or more.

[0019] The cultivation of lactic acid bacteria, the maintenance of bacterial cells, etc. can be carried out by various techniques. For example, as a medium suitable for culturing lactic acid bacteria, a skim milk powder medium can be mentioned, or a liquid medium containing yeast extract, peptone, meat extract, salts, minerals, etc. can be mentioned. Commercially available media include "MRS Broth MERCK" (trade name, Chemicals), "Difco Lactobacilli MRS Broth" (trade name, Nippon Becton Dickinson Co., Ltd.), etc., and such commercially available media may be used.

[0020] The cultivation of lactic acid bacteria is not limited, but for example, it can be carried out statically. Also, in order to suppress the decrease in pH due to metabolites (such as lactic acid) of lactic acid bacteria, the culture (neutralization culture) may be carried out while adding an alkaline agent to the medium to adjust the pH. In that case, as the alkaline agent to be added, for example, an aqueous solution of sodium hydroxide, potassium hydroxide, calcium hydroxide, etc., or ammonia can be used. The pH in the culture may be in the range of pH 5.0 to 7.5, or may be in the range of pH 6.0 to 7.0, and it is preferable to adjust and maintain the pH at that value. The pH adjustment may be performed manually, but it is simple and accurate to use a pH automatic control device (pH stat), etc.

[0021] In the present invention, for lactic acid bacteria, a cell concentrate in which the bacterial cells are concentrated from the state of the culture solution may be prepared, or preferably, it may be freeze-dried after adding an excipient. The cell concentrate can be prepared by concentrating the culture solution as it is, but preferably, the bacteria are collected by means such as centrifugation or filtration, the bacterial cells are further washed with purified water or the like, and suspended in purified water or the like so as to have a predetermined cell concentration. The content of lactic acid bacteria cells in 100% by mass of the cell concentrate may be in the range of 0.1 to 30% by mass in terms of dry bacterial cells, may be in the range of 0.5 to 20% by mass, or may be in the range of 1 to 10% by mass.

[0022] The cell concentrate may also contain an excipient. According to this, even after freezing or freeze-drying and after reconstitution with water, the properties as live bacteria are likely to be maintained. Also, as another aspect, when the cells are crushed and dispersed, re-aggregation of the resulting lactic acid bacteria powder can be prevented. The content of the excipient may be in the range of 10 to 99% by mass in terms of dry matter, may be in the range of 20 to 95% by mass, and may be in the range of 50 to 90% by mass.

[0023] The excipient is not particularly limited, and examples thereof include dextrin; maltodextrin; xanthan gum; sugar alcohols such as lactitol, maltitol, mannitol, sorbitol, xylitol; saccharides such as dextrose, fructose, glucose, lactose, sucrose; organic acids such as adipic acid, citric acid, fumaric acid, glutaric acid, malic acid, succinic acid, tartaric acid, and the like.

[0024] The lactic acid bacteria may be subjected to a treatment of crushing and dispersing. For example, the cell concentrate can be crushed and dispersed using means such as stirring, a mixer, a homogenizer, a ball mill, a bead mill, a jet mill, a generator, and the like.

[0025] The lactic acid bacteria used in the present invention are lactic acid bacteria that can be prepared as described above, and dead cells thereof are used. For example, before or after the step of crushing and dispersing the cell concentrate, the cell concentrate can be heat-treated. Further, after the step of crushing and dispersing the cell concentrate, it can be dried into a powder. Examples of the dry powdering method include freeze-drying, vacuum spray drying, spray drying using hot air, and the like. In addition, by performing spray drying using hot air (spray dry), usually, the lactic acid bacteria are lost and dead cells can be obtained.

[0026] On the one hand, the dietary fiber used in the present invention is not particularly limited. For example, it includes high amylose corn starch, pectin, chitin, glucomannan, alginic acid, β-glucan, inulin, resistant dextrin, carrageenan, fucoidan, guar gum, xanthan gum, gellan gum, pectin, sodium alginate, crystalline cellulose, etc. Particularly preferred is high amylose corn starch.

[0027] As the high amylose corn starch, for example, commercially available products such as "Amylofiber SH" from J-Oil Mills, "Hi-Maize 1043" from Ingredion, "Rodestar" from Nippon Shokuhin Kako, "Amylogel HB-450" from Sanwa Starch Industry, etc. may be used. Also, for example, as shown in JP-A-10-195104 and WO2008 / 155892, those obtained by subjecting to heat treatment or the like to increase the dietary fiber content can also be used. For example, it is preferable to use those in which the dietary fiber content is increased to 40% by mass or more, more preferably those increased to 50% by mass or more, and even more preferably those increased to 60% by mass or more. The dietary fiber content can be measured by the "enzymatic-gravimetric method" or "enzymatic-HPLC method" described in the Food Labeling Law.

[0028] On the other hand, the resistant oligosaccharide used in the present invention is not particularly limited. For example, it includes galactooligosaccharide, fructooligosaccharide, soy oligosaccharide, raffinose, isomaltooligosaccharide, etc. Particularly preferred is galactooligosaccharide.

[0029] If it is a water-soluble dietary fiber or a resistant oligosaccharide as exemplified above, it can effectively serve as a nutrient source for intestinal bacteria in the intestine. Therefore, combined with the effect of the above-mentioned dead lactic acid bacteria cells in improving the intestinal flora, the effect of promoting the production of short-chain fatty acids in the intestine is high.

[0030] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention uses, as active ingredients, dead cells of lactic acid bacteria that can be prepared as described above, dietary fiber, and / or indigestible oligosaccharides, and by administering this to humans or animals other than humans, it promotes the production of short-chain fatty acids in the intestine. The administration method is not particularly limited, but for example, it is preferably administered orally. The short-chain fatty acids in the intestine are specifically acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid, lactic acid, succinic acid, etc. The composition for promoting the production of short-chain fatty acids in the intestine of the present invention is considered to increase the concentrations of acetic acid, propionic acid, and / or butyric acid by particularly promoting the production of acetic acid, propionic acid, and / or butyric acid by the intestinal flora in the large intestine.

[0031] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention can obtain the effects of promoting the absorption of minerals by acetic acid and propionic acid and suppressing the onset of colorectal cancer by butyric acid, for example, by increasing the short-chain fatty acids in the intestine (see the following Reference 1). Also, for example, by improving the barrier function of the intestinal tract and acting on regulatory T cells (Tregs) in the intestinal mucosa, excessive immunity can be suppressed, and allergic symptoms and symptoms of infectious enteritis can be alleviated (see the following Reference 2). Also, for example, energy consumption can be increased via GPR41, etc., which is expressed in intestinal L cells and is known as a receptor for short-chain fatty acids (see the following Reference 2). Also, for example, fat accumulation can be suppressed via GPR43, etc., which is also expressed in intestinal L cells and is known as a receptor for short-chain fatty acids (see the following Reference 2).

[0032] ·Reference 1: Hiroshi Hara, "Physiological Effects of Short-Chain Fatty Acids Produced in the Large Intestine from Prebiotics", Journal of Intestinal Bacteriology (2002) 16: pp. 35-42 · Literature 2: Hidenori Shimizu, Ryuji Ohue-Kitano, Ikuo Kimura "Regulation of host energy metabolism by gut microbiota-derived short-chain fatty acids" Glycative Stress Research 2019; 6 (3): 181-191

[0033] In another aspect of the present invention, the composition for promoting the production of short-chain fatty acids in the intestine according to the present invention can also suppress, for example, the proportion of microorganisms belonging to the genus Firmicutes among intestinal bacteria. Alternatively, for example, the proportion of microorganisms belonging to the genus Bacteroidetes among intestinal bacteria can be increased. Thereby, the Firmicutes / Bacteroidetes ratio can be decreased to adjust the balance of the intestinal flora.

[0034] The dosage of the composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be appropriately set according to the health status and age of the subject, or the degree of the effect of promoting the production of short-chain fatty acids in the intestine required. Typically, in terms of the intake amount in terms of the dried product of dead lactic acid bacteria cells, it may be in the range of 0.001 mg to 100 mg / day / kg body weight, may be in the range of 0.01 mg to 20 mg / day / kg body weight, and may be in the range of 0.1 mg to 10 mg / day / kg body weight. Also, in terms of the intake amount in terms of the dried product of dietary fiber and / or indigestible oligosaccharides, it may be in the range of 1 mg to 1000 mg / day / kg body weight, may be in the range of 5 mg to 500 mg / day / kg body weight, and may be in the range of 10 mg to 300 mg / day / kg body weight.

[0035] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be provided as a food or drink for promoting the production of short-chain fatty acids in the intestine. That is, the above-mentioned dead cells of lactic acid bacteria, dietary fiber, and / or indigestible oligosaccharides may be used as they are, or in combination with other raw materials for food and drink, to form a food or drink. Examples of the raw materials for food and drink to be combined with the above-mentioned dead cells of lactic acid bacteria, dietary fiber, and / or indigestible oligosaccharides include, for example, various carbohydrates, emulsifiers, sweeteners, acidulants, fruit juices, flavors, and the like. More specifically, sugars such as glucose, sucrose, fructose, and honey, sugar alcohols such as sorbitol, xylitol, erythritol, lactitol, and palatinite, and emulsifiers such as sucrose fatty acid ester, glycerin sugar fatty acid ester, and lecithin can be mentioned. In addition to these, various vitamins such as vitamin A, vitamin Bs, vitamin C, and vitamin E, herb extracts, cereal components, vegetable components, milk components, and the like can be mentioned.

[0036] Examples of foods and drinks include, for example, confectioneries such as cookies, senbei, jelly, yokan, yogurt, and manjyu, soft drinks, nutritional drinks, soups, and the like, but are not limited thereto. In addition, other examples of foods and drinks include health foods, supplements, foods for specified health use, or foods with functional claims for promoting the production of short-chain fatty acids in the intestine, and may be provided in forms such as tablets, granules, powders, capsules, drinks, and jelly.

[0037] On the other hand, the composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be provided as a material for adding to foods and drinks for promoting the production of short-chain fatty acids in the intestine, which is used as an additive to foods and drinks as described above.

[0038] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be provided as a pharmaceutical for promoting the production of short-chain fatty acids in the intestine. That is, the above-mentioned dead lactic acid bacteria cells, dietary fiber and / or indigestible oligosaccharides may be used as they are, or in combination with other pharmaceutical raw materials to form a pharmaceutical composition. There is no particular limitation on the other pharmaceutical raw materials combined with the above-mentioned dead lactic acid bacteria cells, dietary fiber and / or indigestible oligosaccharides. If necessary, pharmaceutically acceptable bases and carriers may be added, and by a known formulation method, for example, in the form of tablets, granules, capsules, pills, powders, liquids, powders, jelly-like agents, confectionery agents, etc., and this can be used as an oral preparation.

[0039] On the other hand, the composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be a material for adding to a pharmaceutical for promoting the production of short-chain fatty acids in the intestine, which is used as a material added to such a pharmaceutical.

[0040] The composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be an animal feed for promoting the production of short-chain fatty acids in the intestine. That is, for example, the above-mentioned dead lactic acid bacteria cells, dietary fiber and / or indigestible oligosaccharides may be used as they are, or in combination with other animal feed raw materials to form a composition for animal diet. For example, it may be used in animal feeds such as livestock, racehorses, ornamental animals, pets, etc.

[0041] On the other hand, the composition for promoting the production of short-chain fatty acids in the intestine according to the present invention may be a material for adding to an animal feed for promoting the production of short-chain fatty acids in the intestine, which is used as a material added to such an animal feed.

[0042] In the composition for promoting the production of short-chain fatty acids in the intestine according to the present invention, the contents of the above-mentioned dead lactic acid bacteria cells, dietary fiber and / or indigestible oligosaccharides may be appropriately determined in consideration of the relationship between the amount used and the effective dose in various forms. Typically, in terms of the content in terms of the dry matter of the dead lactic acid bacteria cells, it may be in the range of 0.001 to 99.9% by mass, may be in the range of 0.1 to 50% by mass, and may be in the range of 10 to 30% by mass. Also, in terms of the content converted to the number of cells, 1×10 4~1×10 8 cfu / g may be in the range of, 1×10 5 ~5×10 7 cfu / g may be in the range of, 1×10 6 ~3.3×10 7 cfu / g may be in the range. Also, in terms of the content in terms of the dried product of dietary fiber and / or indigestible oligosaccharide, it may be in the range of 0.1 to 99.9% by mass, may be in the range of 1 to 50% by mass, and may be in the range of 10 to 30% by mass.

Examples

[0043] The present invention will be specifically described below with reference to the following examples, but these examples do not limit the scope of the present invention.

[0044] <Test Example 1> As the heat-killed cells of lactic acid bacteria, heat-killed cells of Enterococcus faecalis (trade name "nano-type lactic acid bacteria nEF (Enterococcus faecalis KH2)", manufactured by Bio Research Co., Ltd.) were used. Also, as the dietary fiber, high amylose corn starch (trade name "Hi-Maize 1043", manufactured by Ingredion Japan Co., Ltd.) was used.

[0045] To the feed for breeding rats, as the heat-killed cells of lactic acid bacteria (sometimes referred to as "nEF"), a blending concentration of 0.005% by mass was added, and as high amylose corn starch (sometimes referred to as "HACS"), a blending concentration of 8% by mass was added, respectively.

[0046] As test animals, Wister male rats (9 weeks old) were divided into four groups (n = 6): (1) "Control", a test group without adding the test sample to the feed; (2) "nEF", a test group with only heat-killed lactic acid bacteria added; (3) "HACS", a test group with only high amylose corn starch added; and (4) "HACS + nEF", a test group with both heat-killed lactic acid bacteria and high amylose corn starch added, and each group was bred with each feed for 3 weeks.

[0047] (Measurement of the amount of short-chain fatty acids in cecal contents) Approximately 0.3 g of cecal contents was dissolved in a 90% ethanol solution and stored at -18°C until measurement. After labeling according to the manual of YMC-Pack FA labeling reagent (manufactured by YMC Co., Ltd.), it was analyzed by HPLC. The mobile phase was acetonitrile / methanol / water = 30:16:54 (pH 4.5), the column was YMC-Pack FA column (4.6×250), and detection was performed at 1.2 mL / min and 400 nm.

[0048]

Table 1

[0049] As a result, as shown in Table 1 and Figure 1, when rats were fed high amylose corn starch together with normal feed, there was a tendency for an increase in the concentration of intestinal short-chain fatty acids such as acetic acid, propionic acid, and n-butyric acid. On the other hand, according to the combined intake of heat-killed lactic acid bacteria and high amylose corn starch, the increase was particularly remarkable in n-butyric acid.

[0050] <Test Example 2> As the heat-killed lactic acid bacteria, heat-killed cells of Enterococcus faecalis (trade name "nano-type lactic acid bacteria nEF (Enterococcus faecalis KH2)", manufactured by Bio Research Co., Ltd.) were used as in Test Example 1. In addition, galactooligosaccharide (manufactured by Fujifilm Wako Pure Chemical Corporation) was used as the indigestible oligosaccharide.

[0051] Galactooligosaccharide (sometimes referred to as "GOS") was added to the mouse breeding feed at a blending concentration of 5% by mass, and heat-killed lactic acid bacteria were added at a blending concentration of 0.08% by mass.

[0052] As test animals, BALB / c mice (7 weeks old) were divided into four groups (n = 3): (1) "Control" as a test group without adding the test sample to the feed, (2) "nEF" as a test group adding only dead lactic acid bacteria cells, (3) "GOS" as a test group adding only galactooligosaccharides, and (4) "GOS + nEF" as a test group adding both dead lactic acid bacteria cells and galactooligosaccharides. They were reared on each feed for 3 weeks. The cecum was collected, the weight in the contents was measured, and the concentration of short-chain fatty acids was measured in the same manner as in Test Example 1.

[0053]

Table 2

[0054]

Table 3

[0055] As a result, as shown in Table 2 and Figure 2, when mice were fed galactooligosaccharides together with normal feed, there was a tendency for an increase in the concentration of intestinal short-chain fatty acids such as acetic acid, propionic acid, and n-butyric acid. On the other hand, when taking a combination of dead lactic acid bacteria cells and galactooligosaccharides, the increase was more significant. Also, as shown in Table 3 and Figure 3, it was consistent with the change in the total amount of cecal contents, and it was considered that the amount of short-chain fatty acids increased due to the promotion of intestinal fermentation.

[0056] From the above, it became clear that when dead lactic acid bacteria cells are used in combination with indigestible components, the action effect of promoting the production of short-chain fatty acids in the intestine is significantly enhanced compared to taking them alone.

Claims

1. A composition for promoting the production of short-chain fatty acids in the intestine, characterized by containing dead cells of lactic acid bacteria and dietary fiber and / or indigestible oligosaccharides.

2. The composition for promoting the production of short-chain fatty acids in the intestine according to Claim 1, wherein the dead cells of lactic acid bacteria are dead cells of Enterococcus faecalis.

3. The composition for promoting the production of short-chain fatty acids in the intestine according to Claim 1 or 2, wherein the dietary fiber is resistant starch.

4. The composition for promoting the production of short-chain fatty acids in the intestine according to any one of Claims 1 to 3, which is used to increase the intestinal production amount of one or more short-chain fatty acids selected from the group consisting of at least acetic acid, propionic acid, and butyric acid.

5. The composition for promoting the production of short-chain fatty acids in the intestine according to any one of Claims 1 to 4, which is a food or drink, a raw material for adding to food or drink, a pharmaceutical, a raw material for adding to a pharmaceutical, an animal feed, or a raw material for adding to an animal feed.

6. A food or drink containing the composition for promoting the production of short-chain fatty acids in the intestine according to any one of Claims 1 to 5.

Images