JP4054697B2 - Constipation improving agent - Google Patents

Description

【００４１】
表１から、乳酸菌エンテロコッカス・フェカリス（ Enterococcus faecalis ）菌体のみを投与した場合には、若干の便秘改善効果が認められるが、アウレオバシジウム培養物のみを投与した場合は、便秘改善効果がほとんど認められないことが分かる。一方、アウレオバシジウム培養物と乳酸菌エンテロコッカス・フェカリス（ Enterococcus faecalis ）菌体を併用した場合、排便回数の増加が認められ、明らかに便秘が改善されていることが分かる。
【００４２】
【発明の効果】
以上説明したように、本発明の便秘改善剤は、アウレオバシジウム属（Aureobasidium sp.）に属する菌であるアウレオバシジウム プルランス M-1 （ Aureobasidium pullulans M-1 ）（ FERM P-19213 ）を培養して得られるβ−１，３−１，６−グルカンを含む培養物をそのまま用いているので、該培養物に含まれる様々な有用成分を損なうことなく利用することができる。そして、該培養物に乳酸菌エンテロコッカス・フェカリス（Enterococcusfaecalis）菌体を配合しているので、これらの成分の相乗効果によって優れた便秘改善効果だけでなく、免疫賦活効果等も期待できる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a constipation-improving agent comprising a culture of a bacterium belonging to the genus Aureobasidium containing β-glucan, particularly β-1,3-1,6-glucan, and lactic acid bacteria as active ingredients, and It relates to the food and drink it contains.
[0002]
[Prior art]
Β-1,3-1,6-glucan produced by a bacterium belonging to the genus Aureobasidium sp. (Commonly known as black yeast) has an immunopotentiating action, an antitumor activity, a cancer cell growth inhibitory action, an antiallergy It is known to have various physiological activities such as action, anti-inflammatory action, cholesterol lowering action, antithrombotic action, dietary fiber action, blood pressure lowering action, blood glucose lowering action, liver function enhancement, etc. It is used as.
[0003]
For example, as an example in which the above β-1,3-1,6-glucan is used for food and pharmaceutical use, particularly for the prevention and improvement of constipation, the following Patent Document 1 discloses incomplete fungus Aureobasidium. An intestinal regulating agent and other medicines based on a polysaccharide produced by a bacterium of the genus ( Aureobacidium ) (Deposit No. 4257, Microtechnical Laboratory) are disclosed.
[0004]
Moreover, the following patent document 2 discloses a method for producing a food or drink mainly composed of fructooligosaccharide and β-1,3-1,6 glucan. This food and drink is a health maintenance beverage (intestinal bifidos). It is described that it can be used for fungus growth, prevention of constipation, immune enhancement, intestinal preparation and the like.
[0005]
Patent Document 3 below discloses a composition containing β-1,3-1,6-glucan and an apple extract, and that the composition is useful as a beverage or a skin coating agent. The beverage includes various allergic symptom reduction effects, immune abnormality improvement effects, cancer suppression effects, vascular disorder disease improvement effects, viral disease improvement effects, urological disease improvement effects, constipation, diarrhea, etc. It describes that an improvement effect on digestive system diseases can be expected.
[0006]
[Patent Document 1]
JP-A-57-149301 [Patent Document 2]
Japanese Patent Publication No. 5-4063 [Patent Document 3]
Japanese Patent Laid-Open No. 2002-335926
[Problems to be solved by the invention]
However, the concentration of β-1,3-1,6-glucan contained in the culture solution of the bacterium belonging to the genus Aureobasidium ( Aureobasidium sp. ) Is low (usually around 0.2% (w / v), Since it is 0.5 to 0.6% (w / v) at most, when using the culture solution as it is described in Patent Document 2, it is sufficient if it is not ingested in a relatively large amount There was a problem that no physiological activity could be expected.
[0008]
Further, when purification is performed as described in Patent Document 1 in order to increase the β-1,3-1,6-glucan concentration, not only the purification cost is required but also the culture solution is used in the purification process. There is a problem that other useful components contained therein (for example, phosphorus, potassium, magnesium, vitamin C, oleic acid, linoleic acid, etc., which are components that assist in the absorption of β-glucan) are lost.
[0009]
On the other hand, in Patent Document 3, isomaltoligosaccharide, galactooligosaccharide, xyl sucrose, xylo-oligosaccharide, chitosan, glycomacropeptide for the purpose of complementing the bifido activity in the culture medium of microorganisms belonging to the genus Aureobacidium. It is described that wheat fiber, corn fiber, soybean oligosaccharide, hemicellulose, soybean peptide or the like can be blended, but as described above, since the β-1,3-1,6-glucan content in the culture solution is low, Even when the above components are used in combination, a sufficient effect of improving constipation cannot be expected.
[0010]
Therefore, the object of the present invention is to use various useful components contained in the culture solution of bacteria belonging to the genus Aureobacidium ( Aureobacidium sp. ) Without damaging them, and has an excellent constipation-improving effect. It is in providing the constipation improving agent which can anticipate an effect etc., and the food-drinks containing it.
[0011]
[Means for Solving the Problems]
In order to achieve the above object, the constipation-improving agent of the present invention contains aureobasidium. Pullulans M-1 (Aureobasidium pullulans M- 1) and cultures containing beta-1,3-1,6-glucan obtained by culturing (FERM P-19213), Lactobacillus Enterococcus faecalis (Enterococcus faecalis) cells As an active ingredient.
[0012]
The constipation improving agent of the present invention is aureobasidium. Since a culture containing β-1,3-1,6-glucan obtained by culturing pullulan M-1 ( Aureobasidium pullulans M-1 ) ( FERM P-19213 ) is used as it is, β contained in the culture Various useful components other than -1,3-1,6-glucan can also be used without damage. And, since the mixing lactic acid bacteria Enterococcus faecalis (Enterococcusfaecalis) cells in the culture, by the synergistic effect of these components, as well as excellent constipation improvement can be expected immunopotentiating effects like.
[0013]
In constipation-improving agent of the present invention, since use of the A Ureobashijiumu pullulans M-1 (Aureobasidium pullulans M- 1) (FERM P-19213), to obtain a more bioactive beta-1,3-1,6-glucan be able to.
[0014]
Moreover, it is preferable to contain 1-40 mass% of said cultures in conversion of (beta) -1,3-1,6-glucan and 4-95 mass% of said lactic acid bacteria in solid content.
[0015]
Furthermore, the culture preferably contains 1% by mass or more of β-1,3-1,6-glucan in the solid content.
[0018]
Furthermore, it is preferable that the lactic acid bacteria are heat-sterilized. According to this aspect, a constipation improving agent that can be widely added to foods and drinks that require heat treatment, has high storage stability, and is extremely safe when used as a raw material for foods and drinks or pharmaceuticals. Can be provided.
[0020]
DETAILED DESCRIPTION OF THE INVENTION
In the present invention, Aureobasidium which is a bacterium belonging to the genus Aureobasidium sp. Pullures M-1 ( Aureobasidium pullulans M-1 ; National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center Deposit No. FERM P-19213 ) is used. As a culture obtained by culturing the same (hereinafter simply referred to as a culture), a culture solution itself obtained by culturing a fungus capable of producing β- 1,3-1,6-glucan, or a concentrated solution of the culture solution Alternatively, a solid material obtained by removing moisture from the culture solution can be used.
[0021]
Contact name in the present invention, the beta-1,3-1,6-glucan, means those having a glucose glucose branches in beta-1, 6 bonds from beta-1, 3 linked backbone structure To do.
[0022]
The above Aureobasidium genus (Aureobasidium sp.) Is a bacterium belonging to the Aureobasidium Culture of pullures M-1 ( Aureobasidium pullulans M-1 ) ( FERM P-19213 ) can be carried out according to a known method (see JP-A-57-149301, etc.). That is, a medium (pH 5.2-6.0) containing 0.5 to 1.0% by mass of carbon source (sucrose), 0.1% by mass of N source, and other trace substances (for example, vitamins and inorganic substances). The inoculum is inoculated and cultured at a temperature of 20 to 30 ° C. for 2 to 3 days, preferably aerated and stirred. As β-1,3-1,6-glucan is produced, the viscosity of the culture solution increases and becomes a highly viscous gel. The culture broth thus obtained usually contains 0.6 to 1.8% by mass of solid content, and 5-1, β-1,3-1,6-glucan is contained in the solid content. -80 mass% is contained. In addition to β-1,3-1,6-glucan, for example, other useful components such as phosphorus, potassium, magnesium, vitamin C, oleic acid, linoleic acid, which are components that help absorb glucan are included. Therefore, the physiological activity effect of β-1,3-1,6-glucan can be efficiently exhibited. In the present invention, a culture containing 1% by mass or more of β-1,3-1,6-glucan in the solid content is preferably used, and 5 β-1,3-1,6-glucan is contained in the solid content. A culture containing at least mass% is more preferably used. If the β-1,3-1,6-glucan concentration in the culture is too low, the bioactive effect of the glucan cannot be sufficiently expected.
[0023]
The β-1,3-1,6-glucan can be quantified according to the method described in Japanese Examined Patent Publication No. 3-48201. That is, after completion of the culture, the culture broth was sterilized and centrifuged to remove the cells, and 10% (v / v) chloroform / butanol mixture was added to the resulting solution and shaken (Sevage method). Thereafter, the mixture is centrifuged to remove chloroform and insoluble matters. After repeating this operation twice, the precipitate is recovered by ethanol precipitation and dissolved in distilled water, the pullulan is decomposed by enzyme treatment, dialyzed in distilled water, and the dialysate is ethanol precipitated to precipitate. The product (β-1,3-1,6-glucan) may be recovered to obtain the yield.
[0024]
In the present invention, the culture solution obtained as described above may be used as it is by sterilization by heating or pressure heating, or may be used after sterilization after separating and removing cells by centrifugation or the like. Moreover, what was concentrated as needed and also what was dried can also be used. In addition, the culture of bacteria belonging to the genus Aureobasidium ( Aureobasidium sp. ) Is used as a food additive such as a thickening stabilizer and has high safety.
[0025]
In the present invention, lactic acid bacteria, use of Lactobacillus Enterococcus faecalis (Enterococcus faecalis). Lactic acid bacteria Enterococcus faecalis (Enterococcus faecalis) may be used alone, Enterococcus faecalis (Enterococcusfaecalis), Enterococcus faecium (Enterococcus faecium), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), Streptococcus・ Cremorris ( Streptococcus cremoris ), Streptococcus lactis , Streptococcus thermophilus ( Streptococcus thermophilus ), Bifidobacterium Longum , Bifidobacterium breveum , Bifidobacterium breve bacterium At least one lactic acid bacterium such as Bifidobacterium bifidum may be used in combination.
[0026]
Incidentally, Enterococcus faecalis (Enterococcus faecalis), Enterococcus faecium (Enterococcus faecium) is a lactic acid bacteria used in the lactobacillus preparation or the like. Lactobacillus casei and Lactobacillus acidophilus are lactic acid bacteria used in cheese, fermented milk, yogurt, lactic acid bacteria beverages and the like. Streptococcus cremoris (Streptococcus cremoris), Streptococcus lactis (Streptococcus lactis), Streptococcus thermophilus (Streptococcus thermophilus) is a lactic acid bacteria used cheese, yogurt and the like. Bifidobacterium longum , Bifidobacterium breve and Bifidobacterium bifidum are lactic acid bacteria used in fermented milk and the like. Therefore, any of these lactic acid bacteria can be easily obtained by those skilled in the art.
[0027]
In the present invention, et Nterokokkasu faecalis (Enterococcus faecalis, for example, ATCC 19433, ATCC 14508, ATCC 123655, IFO 16803 , etc.) can be used. Enterococcus faecalis (Enterococcus faecalis) are known to have a strong immunostimulatory activity, by combination with the above culture, synergistic constipation improvement effect and immunopotentiating effect of these components is expected it can.
[0028]
In the present invention, the lactic acid bacteria are preferably heat-sterilized. Thereby, it can be widely added to foods and drinks that require heat treatment, has high storage stability, and is extremely safe when used as a raw material for foods and drinks and pharmaceuticals.
[0029]
The lactic acid bacteria may be cultured according to a conventional method. For example, cells are collected from a culture obtained by culturing the lactic acid bacteria according to a conventional method by a method such as filtration or centrifugation, washed with water, It is sufficient to suspend it in a heat treatment at 80 to 115 ° C. for 30 minutes to 3 seconds. The heat-sterilized lactic acid bacteria may be used after being concentrated and dried as necessary.
[0030]
The constipation improving agent of the present invention is, for example, Aureobasidium which is a bacterium belonging to the above-mentioned Aureobasidium sp. It can be obtained by mixing and dispersing the heat-sterilized bacterial cells of the lactic acid bacteria in a sterilized culture medium of pullures M-1 ( Aureobasidium pullulans M-1 ) ( FERM P-19213 ) . Moreover, it can also be set as various forms, such as a tablet, a capsule, a powder, a granule, a liquid form, a paste form, and a jelly form, as needed.
[0031]
The constipation-improving agent of the present invention contains 1 to 40% by mass of the culture in terms of β-1,3-1,6-glucan and 4 to 95% by mass of the lactic acid bacteria in the solid content. Preferably, the culture contains 2 to 40% by mass in terms of β-1,3-1,6-glucan, and more preferably contains 10 to 95% by mass of the lactic acid bacteria. It is particularly preferable to contain 3 to 40% by mass of the product in terms of β-1,3-1,6-glucan and 30 to 95% by mass of the lactic acid bacteria. In addition to the above basic components, fragrances, sweeteners, vitamins, minerals, oligosaccharides, thickening polysaccharides, dextrin, and the like can be included as appropriate.
[0032]
The effective intake of the constipation-improving agent of the present invention is 0.01 to 10 g of the culture in terms of β-1,3-1,6-glucan and 0.01 to 10 g of the lactic acid bacteria per adult day. Preferably, the culture is 0.5 to 5 g in terms of β-1,3-1,6-glucan, and the lactic acid bacterial cell is 0.05 to 1 g.
[0033]
Moreover, the constipation improving agent of this invention can also be mix | blended with various food-drinks, such as a soft drink, a jelly drink, fruit juice drink, vegetable juice, soup, miso soup, for example. The amount of the constipation-improving agent in each of the foods and drinks may be set based on the effective daily intake per adult, but is usually preferably 1 to 50% by mass, more preferably 10 to 20% by mass. . In addition, there is no restriction | limiting in particular in the addition method, It can also add from the beginning with the other raw material used for each food-drinks.
[0034]
【Example】
Hereinafter, the present invention will be specifically described with reference to examples. In the following description, “%” represents “mass%” unless otherwise specified.
[0035]
Example (1) Cultivation of Aureobasidium Aureobasidium pullulans M-1 (FERM P-19213) was precultured with 1% sucrose, 0.2% ascorbic acid, and rice bran 0. An appropriate amount was inoculated into a liquid medium (pH 5.3) containing 2%, followed by aeration and agitation culture at 25 ° C. for 2 days. After completion of the culture, this culture solution was sterilized at 121 ° C. for 15 minutes. This culture solution had a solid content of 1% by mass, and contained 35% by mass of β-1,3-1,6-glucan in the solid content.
[0036]
(2) Enterococcus faecalis (Enterococcus faecalis) culturing Enterococcus faecalis (Enterococcus faecalis, IFO 16803) and, 37 ° C. in Rogosa medium, the preculture was incubated for 24 hours, yeast extract 4%, polypeptone 3%, lactose 10 An appropriate amount was inoculated into a liquid medium containing 1%, and neutralization culture was performed at 37 ° C. for 22 to 24 hours while adjusting with a sodium hydroxide aqueous solution to pH 6.8 to 7.0 using a pH stat.
[0037]
After completion of the culture, the cells were separated and collected with a continuous centrifuge, then diluted with water to the original liquid volume, and again separated and collected with a continuous centrifuge. This operation was repeated a total of 4 times to wash the cells. Next, the washed cells were suspended in an appropriate amount of water and sterilized at 100 ° C. for 30 minutes, and then the cells were dried using a spray dryer to prepare a heat-sterilized cell powder.
[0038]
(3) Confirmation of constipation improvement effect Using the aureobasidium culture obtained in (1) above and the lactic acid bacteria obtained in (2) above, a confirmation test of constipation improvement effect was conducted by the following method. It was.
[0039]
The study period is 4 weeks, and 10 volunteers with constipation do not administer anything during the first week (first week), and lactic acid bacteria (200 mg / day) during the next week (second week). ), The next week (3rd week) is administered aureobasidium culture (15 ml / day), and the last week (4th week) is administered aureobasidium culture (15 ml / day). ) And lactic acid bacteria (200 mg / day). Then, the number of defecations in each test sample during one week was checked. The results are shown in Table 1.
[0040]
[Table 1]

[0041]
From Table 1, when administered alone Lactobacillus Enterococcus faecalis (Enterococcus faecalis) cells, although a slight constipation improvement effect is observed, when administered alone Aureobasidium culture, observed almost constipation improvement I can't understand. On the other hand, when used in combination with Aureobasidium culture lactic acid bacteria Enterococcus faecalis (Enterococcus faecalis) cells, an increase in stool frequency was observed, it can be seen clearly constipation has been improved.
[0042]
【The invention's effect】
As described above, the constipation improving agent of the present invention is Aureobasidium which is a bacterium belonging to the genus Aureobasidium sp. Since a culture containing β-1,3-1,6-glucan obtained by culturing pullulan M-1 ( Aureobasidium pullulans M-1 ) ( FERM P-19213 ) is used as it is, it is contained in the culture. Can be utilized without impairing various useful ingredients. And since the lactic acid bacterium Enterococcus faecalis cell body is mix | blended with this culture, not only the constipation improvement effect excellent by the synergistic effect of these components but the immunostimulation effect etc. can be anticipated.

Claims (4)

Aureobasidium Pullulans M-1 (Aureobasidium pullulans M- 1) and cultures containing beta-1,3-1,6-glucan obtained by culturing (FERM P-19213), Lactobacillus Enterococcus faecalis (Enterococcus faecalis) cells And constipation-improving agent, characterized by comprising During solids, the culture containing 1 to 40% by mass beta-1,3-1,6-glucan conversion, and contains the lactic acid bacteria 4-95 wt%, of claim 1 Constipation improving agent. The constipation improving agent according to claim 1 or 2 , wherein the culture contains 1% by mass or more of β-1,3-1,6-glucan in a solid content. The constipation improving agent according to any one of claims 1 to 3 , wherein the lactic acid bacteria are heat-sterilized.