JP2017171590A - Hematopoietic function promoting composition, and anemia preventive and/or improving compositions - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide food, pharmaceutical, and animal feed compositions and the like useful for promoting hematopoietic function or for preventing and/or improving anemia.SOLUTION: A culture composition of microorganisms belonging to Aureobasidium sp. is used as an active ingredient of a composition used for promoting hematopoietic function or preventing and/or improving anemia. The culture composition of microorganisms belonging to Aureobasidium sp. is also used for producing the composition used for promoting hematopoietic function or preventing and/or improving anemia. This composition is suitably used for foods, pharmaceuticals, animal feed, and the like.SELECTED DRAWING: None

Description

  The present invention relates to a composition useful for food, medicine, animal feed and the like useful for promoting hematopoietic function or preventing and / or improving anemia.

  Aureobasidium pullulans, also known as black yeast, secretes abundant useful components such as β-1,3-1,6-glucan during its culture. Therefore, for example, in the following Patent Document 1, etc., rheumatoid arthritis, collagen disease, emerging autoimmune disease, virus using an aureobasidium culture solution mainly composed of β-1,3-1,6-glucan as an active ingredient・ Bacterial infection, various cancers, cancer invasion, metastasis, carcinogenic virus, neurological disease, dementia, Alzheimer's disease, liver disease, cirrhosis, viral liver disease, intractable disease of unknown cause, lifestyle-related diseases (diabetes, hypertension, arteries) It has been proposed to be used for the prevention or treatment of various plagues such as sclerosis and osteoporosis) or sputum that is very common in Japanese (internal and external hemorrhoids, hemorrhoids).

Japanese Patent Laid-Open No. 2002-204687

  However, it has not been known to use a culture composition of microorganisms belonging to the genus Aureobasidium sp. For the promotion of hematopoietic function or the prevention and / or improvement of anemia. The object of the present invention is to use a culture composition of microorganisms belonging to the genus Aureobasidium sp. For food, medicine, animal useful for preventing and / or improving hematopoietic function or anemia. The object is to provide a composition for food and the like.

  The inventors of the present invention have intensively studied to achieve the above object and have completed the present invention. That is, the present invention is as follows.

[1] A composition comprising a culture composition of microorganisms belonging to the genus Aureobasidium sp. As an active ingredient, and used for promoting hematopoietic function.
[2] A composition comprising a culture composition of a microorganism belonging to the genus Aureobasidium sp. As an active ingredient and used for the prevention and / or improvement of anemia.
[3] The composition of the above-mentioned [1] or [2], which is for foods, pharmaceuticals, or animal feeds.
[4] The composition according to any one of [1] to [3], further comprising an auxiliary active ingredient for preventing and / or improving anemia.
[5] Use of a culture composition of microorganisms belonging to the genus Aureobasidium sp. For the production of a composition used for promoting hematopoietic function.
[6] Use of a culture composition of a microorganism belonging to the genus Aureobasidium sp. For the production of a composition used for the prevention and / or improvement of anemia.
[7] The use according to [5] or [6] above, which is used for production of a composition for food, medicine, or animal feed.
[8] The use according to any one of [5] to [7] above, wherein the composition contains an auxiliary active ingredient for preventing and / or improving anemia.

  ADVANTAGE OF THE INVENTION According to this invention, using the culture composition of the microorganisms which belong to Aureobasidium genus (Aureobasidium sp.), It is useful for food use, a pharmaceutical use, an animal useful for preventing and / or improving an angiogenesis function or anemia. Compositions for food and the like can be provided.

FIG. 1 is a chart showing the results of Test Example 1, and is a chart showing changes in red blood cell counts after 2 months and 4 months of ingestion of Aureobasidium culture solution. FIG. 2 is a chart showing the results of Test Example 1, and is a chart showing changes in the amount of hemoglobin after ingestion of Aureobasidium culture solution after 2 months and 4 months. FIG. 3 is a chart showing the results of Test Example 1, and is a chart showing changes in hematocrit values after 2 months and 4 months of ingestion of Aureobasidium culture solution. FIG. 4 is a chart showing the results of Test Example 1, and is a chart showing changes in MCV (average red blood cell volume) after 2 months and 4 months of ingestion of Aureobasidium culture solution. FIG. 5 is a chart showing the results of Test Example 1, and is a chart showing changes in MCH (average erythrocyte hemoglobin amount) after 2 months and 4 months of ingestion of Aureobasidium culture solution. FIG. 6 is a chart showing the results of Test Example 1, and is a chart showing changes in MCHC (average erythrocyte hemoglobin concentration) after 2 months and 4 months of ingestion of Aureobasidium culture solution.

  In the present invention, a culture composition of a microorganism belonging to the genus Aureobasidium sp. Is used as an active ingredient of a composition used for promoting hematopoietic function or preventing and / or improving anemia. Moreover, the culture composition of microorganisms belonging to the genus Aureobasidium (Aureobasidium sp.) Is used for the production of a composition used for the prevention and / or improvement of hematopoietic function or anemia. This culture composition is rich in β-glucan and other useful components produced by microorganisms belonging to the genus Aureobasidium sp.

  The culture composition of microorganisms belonging to the genus Aureobasidium (Aureobasidium sp.) Used in the present invention (hereinafter also referred to as “Aureobasidium-derived culture composition”) belongs to the genus Aureobasidium sp. Examples include a culture in which microorganisms are cultured, a culture solution in which cells are separated and removed by centrifugation, a concentrated solution of the culture solution, a diluted solution of the culture solution, or a solid product obtained by removing moisture from the culture solution. . In addition to these, a culture composition in which the content of β-glucan is increased by removing low-molecular impurities by ultrafiltration or hydrous ethanol precipitation or by other known fractionation means is also included. It is. The culture of Aureobasidium pullulans is used as a food additive such as a thickening stabilizer and has high safety.

  The above-mentioned aureobasidium-derived culture composition used in the present invention contains β-glucan produced by culturing a microorganism belonging to the genus Aureobasidium sp. In a content of 100 g of the culture. In terms of conversion, it is preferable to contain 50 to 3,000 mg, more preferably 100 to 2,000 mg.

In addition, determination of content of (beta) -glucan can be performed by the following methods, for example. That is, the culture solution is subjected to an enzyme treatment using amylase, amyloglucosidase, protease, or the like to remove proteins and α-glucan such as pullulan, and ethanol precipitation is performed. Further, it is filtered through a glass filter to obtain a polymer sample. At this time, in order to remove low-molecular substances containing monosaccharides, the cells are thoroughly washed with 80% ethanol. The washed polymer sample is further washed with acetone, and sulfuric acid is added for hydrolysis. After hydrolysis, the solution is neutralized, the filtrate is collected, glucose is quantified by the glucose oxidase method, and the value calculated based on the following Equation 1 is defined as the amount of β-glucan.
Formula 1: β-glucan (g / 100 g) = glucose (g / 100 g) × 0.9

  The content of β-glucan can also be determined as the so-called sugar chain-containing polymer substance (polysaccharide) content. In this case, the culture solution is subjected to enzyme treatment using amylase, amyloglucosidase, protease, etc., and α-glucan such as protein and pullulan is removed, followed by ethanol precipitation. Further, it is filtered through a glass filter to obtain a polymer sample. At this time, in order to remove low-molecular substances containing monosaccharides, the cells are thoroughly washed with 80% ethanol. The washed polymer sample is further washed with acetone, and the weight of the polymer sample is measured to determine the amount of the sugar chain-containing polymer substance (polysaccharide).

  Note that β-glucan quantified in this way includes those having functional groups such as sulfate groups and phosphate groups, and polysaccharides composed of sugars other than glucose, which are constituent sugars of β-glucan. Quantified. Therefore, when β-glucan is quantified in this way as a broadly defined sugar chain-containing polymer (polysaccharide), β-glucan produced by culturing microorganisms belonging to the genus Aureobasidium sp. The content of is preferably 70 to 5,000 mg, more preferably 140 to 3,000 mg, in terms of the content with respect to 100 g of the culture mass.

  Examples of microorganisms belonging to the genus Aureobasidium sp. Include Aureobasidium pullulans. More preferably, Aureobasidium pullulans M-1 (Aureobasidium pullulans M-1; National Institute of Advanced Industrial Science and Technology patent biological deposit center deposit number FERM BP-08615) or Aureobasidium pullulans M-2 (Aureobasidium pullulans M-2) pullulans M-2, National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center Accession Number FERM BP-10014). In addition, β-glucan produced by these strains is obtained from the main chain in which glucose is β-1,3 linked by structural analysis by NMR measurement (13C NMR: Varian's UNITY INOVA500 type, 1HNMR: Varian's UNITY INOVA600 type). It has been clarified that it is β-1,3-1,6-glucan having a structure in which glucose is branched by -1,6 bonds.

  Culture of microorganisms belonging to the genus Aureobasidium sp. Can be performed according to a known method (see JP-A-57-149301, etc.). That is, a medium (pH 5.2-6) containing 0.5 to 5.0% by mass of carbon source (sucrose), 0.1 to 5.0% by mass of N source, and other trace substances (for example, vitamins and inorganic substances). 0.0) is inoculated with bacteria, and aeration culture is performed at a temperature of 20 to 30 ° C. for 2 to 14 days, preferably aeration and agitation culture. As β-glucan is produced, the viscosity of the culture solution increases and becomes a highly viscous gel. The thus obtained culture broth usually contains 0.6 to 10% by mass of solid content, and 5 to 80% by mass of β-glucan is contained in the solid content. In addition to β-glucan, other useful components such as phosphorus, potassium, magnesium and vitamin C are also included.

  Culture of microorganisms belonging to the genus Aureobasidium sp. May be performed by adding killed microorganisms to the culture medium for culturing them and assimilating them as nutrient components. In this case, the amount of dead cells in the culture solution is usually about 100 cells / mL (culture solution) to 100 trillion cells / mL (culture solution), although it depends on the type of cells used. Is preferred. Microbial killing can be performed by heat sterilization or the like.

  The microorganism used as the killed microorganism is not particularly limited as long as it contains an aureobasidium microorganism nutrient source such as a nitrogen source, and examples thereof include lactic acid bacteria and yeast. Examples of lactic acid bacteria include Enterococcus faecalis, Enterococcus faecalis, E.fecium, Lactobacillus acidphilus, L. gasseri, L. mali, Lactobacillus plantarum (L.plantarum), Lactobacillus buchneri (L.buchneri), Lactobacillus casei (L.casei), Lactobacillus johnsonii (L.johnsonii), Lactobacillus gallinarum (L.gallinarum) ), Lactobacillus amylovorus (L.amylovorus), Lactobacillus brevis (L.brevis), Lactobacillus rhamnosus (L.rhamnosus), Lactobacillus kefir (L.kefir), Lactobacillus paracasei (L.paracasei) ), Lactobacillus genus such as L. crispatus Bacteria, Streptococcus thermophilus and other Streptococcus bacteria, Lactococcus lactis and other Lactococcus bacteria, Bifidobacterium bifidum, Bifidobacterium longum (B longum), B. adolescentis, B. infantis, B. breve, Bifido catenuratum (B. breve) Bifidobacterium bacteria such as B.catenulatum), Bacillus bacteria such as Bacillus subtilis and B. coagulans, Clostridium bacteria such as Clostridium butilicum, etc. be able to.

  Yeasts include imperfect fungi, including Aureobasidium pullulans, Saccharomyces cerevisiae, Saccharomyces intemedius, Saccharomyces intemedius, Saccharomyces validis (Saccharomyces ellipsoideus), Saccharomyces mali risler (Saccharomyces mali risler), Saccharomyces mandschuricus, Saccharomyces y scharm, Sces・ Saccharomyces piriformis, Saccharomyces Anamensis (Saccharomyce) s anamensis), Saccharomyces cartilaginosus (Saccharomyces cartilaginosus), Saccharomyces Batatae, Saccharomyces Batuse, Saccharomyces Coreusus robustus, Saccharomyces Coreusus robustus Bergensis (Saccharomyces Carlsbergensis), Saccharomyces Monacensis (Saccharomyces Monacensis), Saccharomyces Marxianus, Zygosaccharomyces (Chigocaccharomyces Saccharomyces), Mayor (Zygosaccharomyces ), Hansenula anomala, etc. can be used .

  Moreover, when using the culture composition which raised content of (beta) -glucan as said Aureobasidium origin culture composition, for example, the culture of the microorganisms which belong to Aureobasidium sp. (Aureobasidium sp.) Is filtered with diatomaceous earth etc. After removing the insoluble matter, the low molecular weight compound is adsorbed and removed using powdered activated carbon or the like. What was obtained by performing ultrafiltration with an ultrafiltration membrane and precipitating β-glucan recovered in the non-passing fraction with water-containing ethanol can be used. The culture composition having an increased β-glucan content thus obtained typically has a value obtained by quantifying β-glucan as the above-mentioned broadly-defined sugar chain-containing polymer substance (polysaccharide). , Β-glucan in a solid content of 40 to 100% by mass, and more typically β-glucan in a solid content of 70 to 100% by mass.

  As shown in the Examples described later, the culture composition of a microorganism culture belonging to the genus Aureobasidium sp. Has an effect of enhancing the hematopoietic function. Therefore, for example, a human who wants to prevent anemia or to improve the symptoms can take it by ingesting the aureobasidium-derived culture composition. Anemia includes the following types, causes, and symptoms.

Type: Cause ・ Iron deficiency anemia: Production of red blood cells decreases due to lack of iron, protein, vitamin B12, folic acid, etc.

  • Hemorrhagic anemia: due to continuous bleeding. For example, bleeding due to cancer of the gastrointestinal tract such as the stomach and intestines, bleeding due to uterine fibroids in the case of women, menstruation can also cause bleeding anemia.

  -Hemolytic anemia: The lifespan of erythrocytes is about 90 days, but due to some reason the lifespan is shortened or destruction is excessive beyond the production capacity. There are congenital and acquired. Congenital is due to genetic predisposition. The cause of acquiredness is not well understood.

  -Aplastic anemia: The hematopoietic stem cells themselves in the bone marrow are reduced for some reason. Therefore, not only erythrocytes but also all types of blood cells such as leukocytes and platelets are decreased. There are congenital and acquired, and congenital aplastic anemia is an autosomal recessive inheritance. Acquired aplastic anemia is further classified as idiopathic or secondary.

  Symptoms of anemia include dizziness, dizziness, palpitation / shortness of breath, fatigue / fatigue / weakness, loss of appetite, hair loss / split hair, rough skin, stomatitis / mouth hornitis, tinnitus, dysphagia, discoloration / deformation of nails, Examples include taste changes and dysphagia.

  Furthermore, as shown in the examples described later, the culture composition of a microorganism culture belonging to the genus Aureobasidium sp. Is prominent in hematopoietic function, particularly in MCH (average erythrocyte hemoglobin content). There is an effect of increasing MCV (average red blood cell volume) without giving any change. Therefore, in a preferred embodiment of the present invention, it can be suitably applied particularly to a human diagnosed with microcytic anemia or a human suspected of having iron deficiency anemia or secondary anemia.

  The dose of the aureobasidium-derived culture composition can be appropriately determined depending on the intensity of symptoms, physical condition, age, administration method, frequency of administration, administration timing, and the like. In addition, there are few side effects and long-term intake is possible. Therefore, it is preferable to ingest for a long time and to exert the effect continuously. For example, a typical dose is approximately 0.02 to 200 mg / kg (body weight) per day in terms of the amount of β-glucan, which is one of the contents of the aureobasidium-derived culture composition. Take in amounts. Furthermore, it can be applied to animals such as pets and livestock as well as humans.

  The composition of the present invention only needs to be a composition that allows the above-mentioned aureobasidium-derived culture composition to be taken in an effective amount for the prevention and / or improvement of hematopoietic function or anemia. There is no limit. The culture itself may be used, or it may be formed into a form such as powder, granule, liquid, jelly, or cream by a known method. In addition, additives, excipients, disintegrants, binders, lubricants, surfactants, fats and oils, alcohols, sugar alcohols, water, oligosaccharides, water-soluble polymer polysaccharides, etc. Formulation bases and the like can be added. Furthermore, a protein source, a fat source, a carbohydrate source, a fiber source, vitamins, minerals, sweeteners, flavoring agents, acidulants, flavorings, coloring agents, preservatives, and the like may be blended as necessary. Alternatively, soybean meal, soybean oil, wheat, corn, rice, bran, defatted rice meal and the like suitable for animal feed may be blended.

  In the composition of the present invention, the aureobasidium-derived culture composition is converted into the amount of β-glucan produced by the microorganism belonging to the genus Aureobasidium sp. And it is preferable to contain 0.5-30 mg / g, and it is more preferable to contain 1-20 mg / g. Further, it may be used in combination with or in combination with auxiliary active ingredients useful for the prevention of anemia and improvement of symptoms thereof, for example, iron, folic acid, vitamin B12 and the like. In that case, the supplementary active ingredient is 0.5 to 30 mg / g in the case of iron, 0.5 to 30 mg / g in the case of folic acid, vitamin B12, together with the aureobasidium-derived culture composition in the composition of the present invention. It is preferable to use 0.5-30 mg / g in the case of iron, 1-20 mg / g in the case of iron, 1-20 mg / g in the case of folic acid, and 1-20 mg / g in the case of vitamin B12. It is more preferable to use them.

  Further, in a preferred embodiment of the present invention, as the aureobasidium-derived culture composition, the cells are separated and removed by centrifuging or the like from a culture in which a microorganism belonging to the genus Aureobasidium (Aureobasidium sp.) Is cultured, and sterilized. The treated culture solution contains 0.5 to 30 g in an individual package such as an aluminum vapor deposited film, and further 1 to 20 mg in the case of iron, 1 to 20 mg in the case of folic acid, and 1 to 2 in the case of vitamin B12 with respect to 1 g of the culture solution. A form in which 20 mg is contained is preferable, and 2 to 10 mg in the case of iron, 2 to 10 mg in the case of folic acid, and 2 to 10 mg in the case of vitamin B12 are more preferable.

  The composition of the present invention is typically a pharmaceutical, quasi-drug, functional food, dietary supplement, supplement, health food, veterinary drug, veterinary drug, functional animal food, animal, for example. It can be used in various product forms such as dietary supplements, animal supplements, and animal health foods. Or it can be used in combination with these products. Moreover, you may use it in combination with various food / beverage products.

  EXAMPLES Hereinafter, the present invention will be specifically described with reference to examples, but these examples do not limit the scope of the present invention.

<Preparation Example 1>
A culture solution of Aureobasidium pullulans M-2 (FERM BP-10014) (hereinafter referred to as “Aureobasidium culture solution”) was prepared as follows.

  A pre-prepared preculture solution is inoculated in an appropriate amount into a liquid medium (pH 5.3) containing 1% by mass of sucrose, 0.1% by mass of ascorbic acid and 0.1% by mass of rice bran, and 25 ° C., 72 to 96 hours. Aeration and agitation culture were performed (depending on the production batch). After completion of the culture, this culture solution was sterilized at 121 ° C. for 15 minutes. In addition, the solid content of the obtained culture solution after sterilization was approximately 1.2% by mass.

<Test Example 1>
Ten healthy adults in their 20s were orally ingested 90 g of aureobasidium culture medium a day. Blood samples are collected before ingestion, 2 and 4 months after ingestion, and red blood cell count, hemoglobin content, hematocrit value, MCV (mean erythrocyte volume), MCH (mean erythrocyte hemoglobin content), and MCHC (mean erythrocyte hemoglobin concentration) are measured. went.

  As a result, after 2 months and 4 months from the start of intake, no significant change was observed in the number of red blood cells (Fig. 1), and a slight increase in hematocrit was observed. It was not recognized as significant (FIG. 2).

  On the other hand, regarding MCV (mean erythrocyte volume), a significant increase was observed in both male and female groups (FIG. 3). Note that no significant change was observed in MCH (average erythrocyte hemoglobin content) (FIG. 4), and MCV (average erythrocyte volume) increased therein (FIG. 3), so MCHC (average erythrocyte hemoglobin concentration) was calculated. However, MCHC is a test item that has clinical significance only when anemia is diagnosed, and the decrease in healthy individuals is not considered to indicate anemia or a decrease in hematopoietic function. It was.

From the above results, it was considered that the culture solution of Aureobasidium pullulans has the effect of enhancing the hematopoietic function, and its ingestion is useful for the prevention of anemia and the improvement of symptoms.

Claims (8)

  A composition comprising a culture composition of microorganisms belonging to the genus Aureobasidium sp. As an active ingredient, and used for promoting hematopoietic function.   A composition comprising a culture composition of microorganisms belonging to the genus Aureobasidium sp. As an active ingredient and used for the prevention and / or improvement of anemia.   The composition according to claim 1 or 2, wherein the composition is used for food, medicine, or animal feed.   Furthermore, the composition as described in any one of Claims 1-3 containing the auxiliary | assistant active ingredient for prevention and / or improvement of anemia.   Use of a culture composition of a microorganism belonging to the genus Aureobasidium sp. For producing a composition used for promoting hematopoietic function.   Use of a culture composition of a microorganism belonging to the genus Aureobasidium sp. For the production of a composition used for the prevention and / or improvement of anemia.   7. Use according to claim 5 or 6, wherein the use is for the production of a composition for food, medicine or animal feed.   The use according to any one of claims 5 to 7, wherein the composition comprises auxiliary active ingredients for the prevention and / or amelioration of anemia.