JP2006028307A - Beta glucan - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a β-glucan excellent in physiological functions such as antitumor activity and an immunoenhancement action, high in solubility in water, and capable of sufficiently exhibiting the functionality to a living body. <P>SOLUTION: This β-glucan is 1,3, 1,6-β-D-glucan having a weight-average molecular weight of 5,000 or more and less than 500,000, where the content of a glucose residue with β-1,3 linkage without branching is 1-40% of whole glucose residues. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to β-glucan and β-glucan compositions useful as foods, cosmetics, pharmaceuticals and the like.

  β-glucan is contained in large amounts in cell walls of yeast, basidiomycetous mushrooms, molds, lactic acid bacteria, and other microorganisms. In addition, cells that secrete β-glucan outside the cells are also known (see Patent Document 1). The cell walls and extracts of these microorganisms and basidiomycetes, and β-glucan secreted and produced outside the cells have a high immunity-enhancing effect, and are being applied to food and cosmetics. In particular, β-glucan derived from mushrooms Has also been used as a pharmaceutical.

  As strains that secrete and produce β-glucan outside the cells, there are strains of the genus Aureobasidium (Aureobasidium sp.), and culture solutions and purified products have been used as foods and drinks and food additives. The β-glucan produced by this strain has a β-1,3-linked glucose residue as a main chain, and this glucose residue has a β-1,6-linked glucose residue as a side chain (branched chain). , 3,1,6-β-D-glucan is well known (see Non-Patent Document 1 and Non-Patent Document 2). Analysis of the detailed structure of β-glucan molecules is also progressing, and it has been clarified that organic acids, phosphate groups, and sulfonate groups may be bound to these β-glucans. In addition, the binding mode of glucose residues of β-glucan molecules has been studied, and the ratio of β-1,6-bonded side chains to β-1,3-bonded main chains has been analyzed.

  For example, the Aureobasidium pullulans IFO4466 strain, which has long been known as an Aureobasidium strain and has been known as a conserved strain for several decades, also produces β-glucan. When the same strain is cultured by a known culture method, β-glucan having β-1,3 bond as the main chain and β-1,6-bonded side chain is generated, and the structure of β-glucan is β It has been analyzed that the number of side chains of -1,6 bonds is relatively large. Specifically, β-glucan produced by Aureobasidium pullulans IFO4466 has a number of glucose residues having only β-1,3 bonds in the β-glucan molecule, that is, β-1,6-linked branches. The ratio of the number of unreacted glucose residues to the total number of glucose residues involved in the binding of glucose (total amount including branched glucose residues having 1, 3, 1, 6 bonds) is 44.44% It is reported that such highly branched β-glucan has high antitumor activity and immunostimulatory action (see Patent Document 2). However, since this β-glucan is poorly soluble in water, when used in foods, cosmetics, pharmaceuticals, etc., it has not been possible to obtain excellent antitumor activity or immune enhancing action.

  Further, even when strains that produce pullulan are cultured at a low pH (pH 4.5), by controlling the pH strictly between 6.8 and 8.5, A strain capable of producing only β-D-glucan has been reported (see Patent Document 3), and β-glucan produced by the strain is more highly branched than the above and has a number average molecular weight of 500,000 or more. However, since this β-glucan is also poorly soluble in water, when used in foods, cosmetics, pharmaceuticals, etc., it has not been possible to obtain excellent antitumor activity or immune enhancing action.

  As described above, β-glucan derived from microorganisms and basidiomycetes, for example β-glucan produced by Aureobasidium strain, is known as β-glucan having physiological functions such as antitumor activity. The performance is not fully satisfactory, and there are some that do not dissolve in water and do not exhibit physiological functionality, and β-glucan that exhibits excellent physiological functionality has been desired. In particular, β-glucan obtained from Aureobasidium strains, including β-glucan produced by IFO 4466 strain, is highly branched and has been investigated for antitumor activity, but has the disadvantage of a large molecular weight and poor solubility. It is considered that the functionality of the glucan molecule is limited.

In addition, cellulose is a substance classified into β-glucan in a broad sense consisting of only 1,4 bonds, but is insoluble in water and physiological functions such as antitumor activity are not known. Curdlan is a β-glucan consisting only of β-1,3 bonds and has a molecular weight of several hundreds of thousands to several tens of thousands. However, even if the molecular weight is tens of thousands, it is hardly soluble in water. There are no active reports on antitumor activity. On the other hand, the pharmaceutical schizophyllan obtained from Suehirotake is a β-glucan having a β-1,3-linked branch in the β-1,3-linked main chain, and a β-1,3-linked glucose residue having no branch Is approximately 67% (the ratio having 1,6 bond branches is approximately 33%), and the molecular weight is approximately 400,000.
As described above, β-glucan that is excellent in physiological functions such as antitumor activity and immune enhancing action and that sufficiently exhibits its functionality to living bodies is strongly desired.

JP 2004-49013 A Japanese Patent No. 3444624 JP-A-9-56391 Acta Chemica Scandinavia 17, 1351-1356 (1963) Agric. Biol. Chem. 47 (6), 1167-1172 (1983)

  An object of the present invention is to provide a β-glucan that is excellent in physiological functions such as antitumor activity and immune enhancing action, has high solubility in water, and sufficiently exhibits its functionality for a living body. .

  In the study on the creation of β-glucan molecules having high functionality, the present inventors have an important point that β-glucan has excellent solubility in water in order to exhibit excellent functionality. For sex, we found that the abundance ratio of side chains to the main chain and the molecular weight of β-glucan molecules are closely related. That is, in 1,3,1,6-β-D-glucan, the focus is on the ratio of β-1,3-linked glucose residues without side chains to the total glucose residues and the weight average molecular weight of β-glucan. As a result of the narrowing examination, it has been found that β-glucan having a certain range of branching degree and a certain range of molecular weight exhibits excellent functionality, and has led to the present invention.

  That is, the present invention is a 1,3,1,6-β-D-glucan having a weight average molecular weight of 5,000 or more and less than 500,000, and the proportion of β-1,3 linked glucose residues having no branching. The present invention provides a β-glucan that is 1% to 40% of all glucose residues.

The present invention also provides the β-glucan in which the constituent sugar of β-glucan is only glucose or glucose and mannose.
The present invention also provides the β-glucan obtained by culturing the Aureobasidium pulullans ADK-34 (FERM BP-8391) strain.
The present invention also provides a β-glucan composition comprising the β-glucan in an amount of 1% by weight or more based on the total amount of β-glucan.
The present invention also provides the β-glucan composition containing the β-glucan and a glucan derived from a gramineous plant or 1,3,1,4-β-D-glucan.

  The β-glucan and β-glucan composition of the present invention are excellent in physiological functions such as antitumor activity and immune enhancement, have high solubility in water, and have high immunostimulatory activity especially by oral administration. Therefore, it can be applied to foods, cosmetics, pharmaceuticals and the like, and is particularly useful as a preventive or therapeutic agent for human cancer, or as an infectious disease preventive or therapeutic agent for livestock, pet animals, farmed fish and the like.

Hereinafter, the present invention will be described in detail.
The β-glucan of the present invention has a β-1,3-linked glucose residue as the main chain, and a part of the glucose residue has a β-1,6-linked glucose residue as a branch (also referred to as a side chain). The so-called 1,3,1,6-β-D-glucan, and the proportion of β-1,3-bonded glucose residues having no branching is determined by the total glucose residues (β 1 to 40% of the total of the glucose residues having a branch and the glucose residues having no branch in the -1,3-bonded main chain). This ratio is preferably 10% or more and 30%, more preferably 12% or more and 22.5% or less, from the viewpoint of solubility in water and physiological functionality.

  Furthermore, the β-glucan of the present invention has a weight average molecular weight of 5,000 or more and less than 500,000, and is preferably 5,000 or more and 400,000 or less from the viewpoint of solubility in water and physiological functionality.

  In addition, in the β-glucan of the present invention, the constituent sugar is preferably glucose alone or glucose and mannose from the viewpoint of solubility in water and physiological functionality. In the case of glucose and mannose, the ratio of mannose is preferably 0.1 to 15%.

The β-glucan of the present invention is obtained from microorganisms or basidiomycetes, and may be obtained from any microorganism or basidiomycete, but the microorganisms themselves contain a large amount of β-glucan in their cell walls. Therefore, it is preferable to produce the cells by using microbial cells themselves, extracting from microbial cells, or using microorganisms capable of secreting and producing β-glucan outside the cells.
In addition, β-glucan obtained from microorganisms or basidiomycetes may contain β-glucan having a structure other than the β-glucan of the present invention. In that case, the β-glucan of the present invention is separated and purified for use. However, when the content of β-glucan of the present invention is 1% by weight or more based on the total amount of β-glucan, it may be used as it is as a β-glucan composition. The content of the β-glucan of the present invention in the β-glucan composition is preferably 10% by weight or more, more preferably 30% by weight or more with respect to the total amount of β-glucan.

  In addition, β-glucan includes 1,3,1,4-β-D-glucan and can be obtained from gramineous plants. This 1,3,1,4-β-D-glucan or β-glucan derived from a grass family plant also has physiological functions such as lipid metabolism improving action, intestinal regulating action, blood sugar level increase inhibiting action, etc. Use in combination with the β-glucan of the invention in combination with a β-glucan composition (also referred to as a β-glucan complex) is preferred because the physiological functionality of each is enhanced and a synergistic effect is produced. In addition, solubility in water is also improved, which is preferable. In order to obtain this β-glucan composition (β-glucan complex), 1,3,1,4-β-D-glucan or β-glucan derived from the grass family and the β-glucan of the present invention are used in the present invention. The β-glucan content may be 1% by weight or more, preferably 10% by weight or more, more preferably 30% by weight or more based on the total amount of β-glucan.

  The β-glucan of the present invention is preferably obtained from microorganisms, and the cultured cells obtained by inoculating the microorganisms in a growth medium suitable for the microorganisms and growing the cells remain as they are, or the cultured cells are disrupted to obtain the contents. The cultured cell wall residue obtained by removing the substance can be used. In addition, both β-glucan extracted from the cultured cells or the cultured cell wall residue and purified of the extracted β-glucan can be used. In addition, it is more preferable to use β-glucan secreted and produced outside the cells by culturing microorganisms. In this case, the culture solution after completion of the culture is used as it is or is isolated and purified from the culture solution. Β-glucan can be used.

  Among these, when the cultured cells obtained by inoculating microorganisms in each growth medium and growing the cells are used as they are, the cell contents are reduced in physiological functions, soluble in water, etc. Since there is a possibility of causing a decrease in physical properties, it is preferable to use a cultured cell wall residue obtained by crushing the cultured cells and removing the contents, and further, extracted from the cultured cells or the cultured cell wall residues. It is more preferable to use β-glucan as it is or after purification, and it is most preferable to use β-glucan secreted and produced outside the cells together with the culture solution or isolated and purified from the culture solution.

  As microorganisms for obtaining the β-glucan of the present invention, microorganisms conventionally used for food are preferable because of their high safety. That is, yeasts, lactic acid bacteria, natto bacteria, acetic acid bacteria, koji molds, algae such as chlorella and spirulina, filamentous fungi, microorganisms belonging to the genus Aureobasidium, especially microorganisms belonging to the genus Aureobasidium Is preferred. These can use the said microorganisms isolate | separated from the environment (for example, food, soil, indoors, etc.). Moreover, the stock strain or isolate isolate | separated by single bacteria, Furthermore, the mutant strain which performed mutation operation in them by a conventional method can be used. Examples of the mutation operation include, for example, UV irradiation, chemical treatment with nitrosoguanidine, ethidium bromide, ethyl methanesulfonate, sodium nitrite and the like.

Examples of the yeast include yeasts classified into the genus Saccharomyces used in alcoholic brewing and bread making processes such as beer, happoshu, shochu, sake, wine, whiskey, etc., pressed bread yeast, and soy sauce brewing. Examples thereof include yeasts used, yeasts belonging to the genus Candida used for microbial protein production, and the like.
Examples of the lactic acid bacteria include Lactobacillus genus, Bifidobacterium genus, cocci Leuconostoc genus, Pediococcus genus, Streptococcus genus, and Lactococcus genus. ) Lactic bacteria belonging to the genus are usually used, but other genus Enterococcus, Vagococcus, Carnobacterium, Aerococcus, Can be used. Specific Lactobacillus strains include Lactobacillus bulgaricus, L. helveticus, L. acidophilus, L. lactis, L. lactis, Lactobacillus casei (L.casei), Lactobacillus brevis (L.brevis), Lactobacillus plantarum (L.plantarum), Lactobacillus salmon (L.sake), Streptococcus thermophilus (Streptococcus thermophilus), Streptococcus thermophilus (S. lactis), S. cremoris, Bifidobacterium longum, B. bifidum, B. breve, B. Phytobacterium infantis (B.infantis), Leuconostoc cremoris, Leuconostoc L. mesenteroides, L. ocnos, Pediococcus acidilactici, P. cerevisiae, P. cerevisiae, P. One or more of the conventionally used lactic acid bacteria such as .pentosaceus) can be used. These may be used alone or in combination of two or more. In addition, culture of lactic acid bacteria belonging to the genus Bifidobacterium and culture of other lactic acid bacteria may be performed separately, and these may be mixed.

  Other strains of the genus Bacillus genus Bacillus, strains of the genus Acetobactor acetic acid bacteria, strains of the genus Aspergillus genus and Penicillium genus, chlorella and spirulina Known to produce algae, dried chlorella powder, strains of the genus Aureobasidium, which are known to secrete and produce pullulan, and other polysaccharides used as food additives Xanthomonas genus, Aeromonas genus, Azotobactor genus, Alcaligenes genus, Erwinia genus, Enterobactor genus, Sclerotium genus Pseudomonas ) Strains of the genus, Agrobacterium genus, Macrohomopsis genus can be used

The β-glucan of the present invention may be obtained from basidiomycetes.
Since basidiomycetes contain β-glucan in the nuclei of fruit bodies and mycelia assembled in a lump, the β-glucan of the present invention is obtained by finely pulverizing the fruit bodies and microbial nuclei and extracted from the pulverized product. Or a product obtained by purifying β-glucan from the extract. Also, germination of basidiomycetous spores, inoculating mycelium in each growth medium and growing the bacterial cells, or obtained by culturing the cultured cells and removing the contents Cultured cell wall residue can be used. In addition, any β-glucan extracted from the cultured cells or the residue of the cultured cell wall and purified of the extracted β-glucan can be used. It is also possible to utilize β-glucan secreted and produced outside the bacterial cells by culturing basidiomycetes. In this case, the culture solution after completion of the culture can be used as it is or separated and purified from the culture solution. Β-glucan can be used.

  Among these, β-glucan obtained by finely pulverizing fruiting bodies and mycelia, β-glucan extracted from the finely pulverized product, spore and mycelium are inoculated into each growth medium and cultured by growing the bacterial cells. If the cells are used as they are, the contents of the cells may cause a decrease in physiological functions and physical properties such as solubility in water. Therefore, the cells are obtained by crushing the cultured cells and removing the contents. It is preferable to use the cultured cell wall residue obtained, and it is more preferable to use the cultured cell or β-glucan extracted from the cultured cell wall residue as it is or after purification, and to produce secreted cells outside the cells. It is most preferable to use the obtained β-glucan together with the culture solution or separated and purified from the culture solution.

  The basidiomycete for obtaining the β-glucan of the present invention is most preferably a cultivar, but β-glucan from a basidiomycete not subjected to commercial production can also be used in the present invention. Examples of the basidiomycetes include Agaricus blazei, Amigasatake, Agaricus, Ezoharitake, Enokitake, Kanzotake, Agaricus, Kinugasatake, Kuritake, Salmonella, Sakerihitotake, Sangotake, Shitaketake, Maitake, camellia, winter summer grass, sea cucumber, sea bream, sea bream, green swordfish, green moth, bamboo turkey, sardine mushroom, bamboo shoot, bamboo shoot , Matsutake, Mannentake, Mukitake, Murasakishimeji, Yamadoritake, Yamabushitake, Willow Matsutake, etc. That.

  As a method for isolating cultured cell wall residues of the above microorganisms and basidiomycetes as β-glucan, an appropriate amount of solvent is added to the cultured microorganisms, the cultured mycelium, the cultivated mycorrhiza or fruiting body, and self-digestion Alternatively, there is a method of isolating the cultured cell wall residue as β-glucan by destroying a part of the cell wall by adding hydrolase, allowing the contents to flow away, and recovering the residual components. Also, a method of obtaining β-glucan by damaging microorganisms or basidiomycetous cells by physical force such as a French press or ultrasonic crusher, destroying part of the cells, removing the contents, and collecting the residue There is also.

  Next, a method for extracting β-glucan from microorganisms or basidiomycetes will be described. The method for extracting β-glucan from microorganisms or basidiomycetes is not particularly limited, and extraction may be performed by adding an extraction solvent to microorganisms or basidiomycetes that are the raw materials for extraction. As the extraction solvent, one or two or more mixed solvents such as water, a salt solution, an aqueous acid solution, an aqueous alkaline solution, and an organic solvent can be used. Moreover, extraction efficiency can be improved by using together the enzyme which decomposes | disassembles a cell wall. The extract can be purified by purifying the extract itself in a solid-liquid separation, or a liquid or solid obtained by concentrating β-glucan extracted from the extract by a known method, or purifying the extract by a known method. Any of the liquids, solids, etc. obtained by any production method, any form, and any purity can be used. Of course, there is no problem even if extracted components other than β-glucan are mixed.

  Further, the method for extracting β-glucan from microorganisms or basidiomycetes will be described. The β-glucan of the present invention can be dissolved in a solvent such as water as a water-soluble polymer, and is generally commercially available as basidiomycetes. Mushrooms that have been dried and pulverized with water, warm water, hot water or salt solution, further using acid, alkaline aqueous solution, organic solvent, etc., in a solvent 2 to 100 times the amount of powder for any time It can be extracted at any temperature. Further, β-glucan can be obtained by solid-liquid separation of the extract. Among these, β-glucan extracted with water, warm water or hot water is preferable, and β-glucan extracted with water having a temperature of 80 ° C. or lower and 4 ° C. or higher is more preferable. Further, an extraction accelerator such as an enzyme solution may be added during extraction.

Next, production of β-glucan of the present invention using a microorganism belonging to the genus Aureobasidium will be described.
The β-glucan of the present invention is particularly preferably obtained from a microorganism belonging to the genus Aureobasidium, and is a strain that produces the β-glucan of the present invention inside or outside the cell body by culturing the microorganism. Any of them can be used, and preferable examples thereof are Aureobasidium pullulans strains, specifically, IFO4464, IFO4466, IFO6353, IFO7757, ATCC9348, ATCC3092, ATCC42023, ATCC433032, etc. In particular, Aureobasidium pullulans ADK-34 strain (a strain deposited with the deposit number “FERM BP-8391” at the National Institute of Advanced Industrial Science and Technology (AIST)) is used. The is the most preferred β-glucan yield and isolation of the ease of the present invention. In addition, the microorganisms separated in the environment (for example, food, soil, indoors, etc.) can be used. Moreover, the stock strain or isolate isolate | separated by single bacteria, Furthermore, the mutant strain which performed mutation operation in them by a conventional method can be used. Examples of the mutation operation include, for example, UV irradiation, chemical treatment with nitrosoguanidine, ethidium bromide, ethyl methanesulfonate, sodium nitrite and the like.

  The medium for culturing these microorganisms may be any medium containing nutrient sources (carbon sources, nitrogen sources, inorganic salts) that can be used normally by microorganisms belonging to the genus Aureobasidium, and if necessary, organic A normal medium containing a nutrient can be used, and various synthetic media, semi-synthetic media, natural media, and the like can be used.

  As the carbon source, saccharides are preferable. For example, monosaccharides such as glucose, fructose, mannose, galactose, xylose, arabinose, disaccharides such as sucrose, maltose, lactose, trehalose, oligosaccharides such as fructo-oligosaccharide, xylooligosaccharide, dextrin and starch Polysaccharides can be used alone or in combination. In addition, alcohols such as glycerol and ethylene glycol, sugar alcohols such as mannitol, sorbitol, and erythritol, organic acids, and the like are appropriately used. In addition, beet juice, sugarcane juice, fruit juices including citrus fruits, or those obtained by adding sugar to these juices can be used. The main carbon source is preferably a hexasaccharide such as glucose or fructose, a disaccharide such as sucrose or lactose, a polysaccharide such as starch or dextrin, or a hydrolyzate of these carbohydrates. These carbon sources may be added at any time during the culture. For example, sucrose is added to the medium in a concentration range of 3 to 500 g / l, preferably 5 to 300 g / l, more preferably 10 to 200 g / l. By appropriately feeding as described above, the production rate and production amount of β-glucan can be relatively increased.

  Nitrogen sources include peptone, meat extract, soy flour, casein, amino acids, malt extract, corn steep liquor, casein digest, yeast extract, organic nitrogen sources such as urea, sodium nitrate, ammonium sulfate, ammonia gas, aqueous ammonia, etc. An inorganic nitrogen source or the like can be used alone or in combination.

  As inorganic salts, for example, salts such as sodium chloride, potassium chloride, calcium carbonate, magnesium sulfate, sodium phosphate, potassium phosphate, and cobalt chloride, heavy metal salts, and vitamins can be added and used as necessary. In addition, when foaming occurs during culture, various known antifoaming agents can be appropriately added to the medium.

  The culture conditions for the strain are not particularly limited, and can be appropriately selected within a range in which the strain can grow well. Usually, it is good to culture | cultivate for about 2 to 8 days at pH 5.0-8.5 and 20-35 degreeC. The culture conditions can be appropriately changed according to the type and characteristics of the microorganism strain used, external conditions, etc., and the optimum conditions can be selected. It is preferable to add 1 to 5% (v / v) of the main culture solution as a seed culture solution in the case of flask culture. This does not apply if culture is possible.

  The culture is performed under aerobic conditions such as aeration stirring and shaking. The culture time is preferably performed up to a desired β-glucan production concentration, and is usually about 2 to 5 days. Alternatively, β-glucan may be continuously produced by continuously adding sugars and medium components that are β-glucan substrates. For example, when sucrose is used as the main raw material for β-glucan production, it can be added to the culture solution as a powder or as a liquid sugar dissolved in water at a high concentration. 0.1 to 500 g per liter is preferable. When the addition amount is more than 500 g / l, the production rate is remarkably slow, which is not preferable. After the addition of the substrate, the substrate is preferably treated at 25 to 35 ° C. for 1 to 7 days, particularly about 2 to 5 days, by shaking or aeration and stirring, and the reaction is allowed to proceed under aerobic conditions. Β-glucan can be produced from a certain sucrose.

  The cells obtained by culturing a strain of Aureobasidium pullulans can be isolated, and the β-glucan of the present invention can be produced using the microbial strain or strain extract as a catalyst. In this case, the cultured cells can be used as they are, or, for example, a homogenized cell disruption solution, or the cultured cells or the cell disruption solution can be fixed to an ion exchange resin, ceramic, chitosan, etc. in an alginate gel. What is necessary is just to add to the saccharide | sugar solution which is a substrate, or a culture medium as the immobilized fixed microbial cell. As a solution that can be used for the preparation of a suspension of a cultured bacterial cell, a cultured bacterial cell preparation obtained by crushing or grinding the bacterial cell, or a treated cultured bacterial cell, the above-described medium or Tris-acetic acid, Examples include tris-hydrochloric acid, sodium succinate, sodium citrate, sodium phosphate, potassium phosphate and other buffer solutions alone or in combination. The pH of the buffer is preferably 3.5 to 9.0, more preferably 5.0 to 8.0, and still more preferably 5.2 to 7.8. The amount of cultured cells in the suspension is not limited, but is preferably about 0.1 to 5% in terms of wet volume ratio.

  The addition of saccharides or a medium as a substrate to the suspension can be carried out at any concentration and in any volume, but the amount of substrate added per time is preferably within a range where the activity of the cells can be maintained. For example, 0.1 to 500 g per liter of suspension may be divided into one time or several times, or may be continuously added at about 0.5 to 500 g / day. The solution in which the bacterial cells and the substrate are allowed to act can be incubated at an arbitrary time and temperature to produce β-glucan, but is preferably carried out under the same conditions as the culture of the strain.

  After culturing as described above, β-glucan secreted and produced outside the cells is separated and collected from the culture solution according to a conventional method. Specifically, various known purification means such as separation and removal of solids such as bacterial cells from the culture solution by centrifugation or filtration, and removal of impurities and salts by activated carbon / ion exchange resin are selected and combined. Can be done. For example, adsorption / elution on hydrophobic resin, solvent precipitation using ethanol, methanol, ethyl acetate, n-butanol, etc., column method using silica gel, etc. or thin layer chromatography, preparative high-speed liquid using reverse phase column Separation and purification can be performed by using chromatography alone or in combination as appropriate, and optionally using it repeatedly. Bacterial cells may be sterilized before and after the β-glucan secreted outside the cells is separated by the above method. The sterilization of the bacterial cells may be any temperature as long as the bacterial cells are killed, but is 50 ° C. or higher, preferably 60 ° C. or higher, more preferably 80 ° C. or higher. Further, the secretory β-glucan prepared in the microbial cells can be extracted with hot water by further increasing the temperature, for example, by sterilizing the microbial cells at 90 ° C. or higher, or at 100 ° C. or higher under pressure. it can. The sterilization and hot water extraction time can be set arbitrarily, but it is 10 minutes or more and 8 hours or less, preferably 15 minutes or more and 6 hours or less, more preferably 30 minutes or more and 2 hours or less, and contamination of impurities is suppressed. This is preferable because glucan does not deteriorate.

  The β-glucan and β-glucan composition of the present invention have excellent bioregulatory functionality and physiological activity functions, for example, lipid metabolism improving action, intestinal regulating action, blood sugar level rise inhibiting action, cholesterol lowering action, blood sugar level lowering action, Tumor action, immune enhancement action, immunostimulation action, intestinal immunity enhancement action, skin immunity enhancement action, lifestyle-related disease prevention / amelioration action, etc., excellent solubility in water, emulsification, thickening, etc. And can be preferably used for foods, pharmaceuticals, cosmetics, feeds, chemical products and the like. When used, the β-glucan of the present invention may be used alone, or the β-glucan of the present invention may be used in combination with other components.

  Examples of foods that can be used or blended include oil and fat foods, bakery products, confectionery, noodles, processed rice products, processed wheat products, processed corn products, processed beans (soybeans, red beans, etc.), cereals (buckwheat, crabs, Awa, millet) processed products, agricultural products (potato, sweet potato, taro, troroimo, etc.), dairy products, soups, beverages, seasonings, processed meat products, processed marine products, cooked / prepared foods, health foods, low calories Foods, foods for allergic patients, foods for infants, foods for the elderly, beauty foods, medicinal foods, and the like, and frozen foods, retort foods, instant foods, canned foods and the like thereof can be mentioned.

  Examples of the fat and oil food include margarine, shortening, whipped cream, cream, salad oil, custard cream, dip cream, fat spread, mayonnaise, tartar sauce, dressing, and oiled oil.

  Examples of the bakery product include bread, white bread, confectionery bread, side dish bread, French bread, croissant, pie, castella, sponge cake, butter cake, shoe confectionery, waffle, steamed bread, and fermented confectionery.

  Examples of the confectionery include snacks, donuts, biscuits, crackers, buns, Japanese confectionery, yokan, midway, uiro, dumplings, daifuku mochi, candy, gum, chocolate, strawberry, ice cream, soft cream, sorbet, popsicle , Lact ice, ice confectionery, jelly, pudding, dessert, topping and the like.

Examples of the above processed rice products include cooked rice (including frozen cooked rice, sterile cooked rice, etc.), rice balls, rice ball rice cakes, rolled rice cakes, chili rice cakes, rice cakes, fried rice, pilaf, tea pickles, doria, rice noodles, arabe, rice crackers, etc. Can be mentioned.
Examples of the processed wheat products include cereal, udon, pizza, pasta, hoto, Chinese soba, fried noodles, champon, okonomiyaki, monja-yaki, piroxy, buns, cup noodles and their ingredients.
Examples of the corn processed product include corn flakes, corn snacks, popcorn and the like.
Examples of the processed soybean product include tofu, soy milk, soy milk drink, yuba, deep-fried oil, deep-fried chicken, ganmodoki, red bean paste, miso, various bean dishes, and the like.
Examples of processed cereals (buckwheat, shrimp, abalone, millet), and processed agricultural products (potato, sweet potato, taro, troroimo, etc.) include buckwheat, potato snacks, potato chips, sweet potatoes, potato fries, and various potato dishes. Is mentioned.

Examples of the dairy products include milk, processed milk, yogurt, whey drink, lactic acid bacteria drink, butter, cheese, coffee whitener and the like.
Examples of the soup include potage soup, consomme soup, stew, miso soup, soup, soup, curry and the like, and may be added to these ingredients. Or you may add directly in these like croutons.
Examples of the beverages include soft drinks, carbonated drinks, colas, juices, fruit juices, vegetable juices, tomato juices, shakes, sake, beer, sparkling wine, western liquor, wine, fruit liquors, cocktails, tea, tea, coffee , Cafe au lait, oolong tea, green juice, mineral water, water, etc.
Examples of the above-mentioned seasonings include soy sauce, fish sauce (squid crab soy sauce, sardine soy sauce, salt juice, nanpura, etc.), miso, jam, sauce, toaster sauce, tomato sauce, tomato ketchup, tomato paste, tomato puree, chili sauce, sauce , Pepper, pepper, garlic, ginger, vinegar, chili oil, tabasco, salt, various spices.

Examples of the processed meat product include hamburger, sausage, salami sausage, ham, meatball, meat dumpling, meat bun, dumpling, shumai, and various meat dishes.
Examples of the processed fishery products include kamaboko, deep-fried sweet potato, tsumire, and paste products.
Examples of the health food or medicinal food include supplements, tablets, drinks, sports drinks and the like.

  Examples of pharmaceuticals that can be used or formulated include lipid metabolism improving action, intestinal regulating action, blood sugar level increase inhibiting action, cholesterol lowering action, blood sugar level lowering action, antitumor action, immune enhancing action, immunostimulating action, intestinal tract immune enhancing Medicine, skin immunity enhancing action, lifestyle-related disease prevention / amelioration medicine, cancer prevention or treatment medicine, infection prevention or treatment medicine, skin treatment medicine, ointment, patch, nasal preparation, Examples include ear drops and eye drops. Furthermore, it is also useful as an ointment base, tablet, granule, excipient for powder formulation, internal use, and liquid dispersant.

  Examples of feeds that can be used or formulated include livestock feeds, pet feeds, fish and shellfish feeds, farmed fish feeds, livestock therapeutics or preventives, pet therapeutics or preventives, and seafood therapeutics. Alternatively, prophylactic drugs, cultured fish therapeutic drugs or preventive drugs, infectious disease preventive drugs and the like can be mentioned.

  When the β-glucan and β-glucan composition of the present invention is used for foods, pharmaceuticals, feeds, etc., a substance that gives some physiological activity to the body when ingested may be further blended. For example, cholesterol elevation inhibitor, blood pressure elevation suppressor, blood cholesterol regulation function agent, blood glucose level elevation inhibitor, gut microbiota ameliorator, intestinal regulator, immune enhancer, anticancer agent, antiallergy Examples include agents, digestion / absorption control agents, anti-aging agents, antioxidants, blood circulation promoters, amino acids, peptides, lipids, polysaccharides, oligosaccharides, proteins, carbohydrates, dietary fiber, and enzyme components.

  Specific examples of these substances that give physiological activity include polyunsaturated fatty acids (EPA, DHA) that optimize blood lipid concentration, plant sterols and their esterified substances that regulate serum cholesterol, diacylglycerol, and γ-linolene. Acid, α-linolenic acid, linoleic acid, conjugated linoleic acid and other unsaturated fatty acids, beet fiber, corn fiber, psyllium seed coat, tea polyphenol, lecithin, bonito peptide, sardine peptide, casein deca peptide, soy protein isolate Such as foods and pharmaceuticals containing lactic acid bacteria, gluconic acid, oligosaccharides, various dietary fibers, etc., which improve the intestinal environment and work for intestinal regulation. As other ingredients known to have health functions, chlorella, spirulina, propolis, chitin, chitosan, deoxyribonucleic acid, ribonucleic acid, ganoderma, agaricus, ginkgo biloba extract, citrus fruit , Turmeric, garcinia, apple fiber, gymnema, collagen, blueberry, aloe, saw palmetto, capsanthin, lutein, β-cryptoxanthin, renin, taurine, casein, collagen, glucosamine, casein phosphopeptide (CPP), milk basic protein (MBP) , Lactoferrin, glutathione, theanine, gabba, aspartame, xylitol, recardent, alginic acid, sodium alginate, fucoidan, chondroitin, hyaluronic acid, cellulose, pectin, indigestible Xistrin, glucomannan, inulin, fructan, cyclofructan, difructose, levan, mucin, flavonoid, polyphenol, catechin, tannin, anthocyanin, rutin, quercetin, soy isoflavone, soy saponin, soy globulin, chlorogenic acid, citric acid, lactic acid, Malic acid, capsaicin, sesame lignan, allicin, caffeine, chlorophyll, nattokinase, β-lactoglobulin, plant fermentation enzyme, mevalonic acid, chlorophyll, royal jelly, ginseng, prunes, chamomile, thyme, sage, peppermint, lemon balm, mallow, Herbs such as oregano, catnip tea, Yarrow, Hypiscus, Echinesia, vitamins, calcium fortifiers such as calcium-containing compounds, iron such as iron-containing compounds Reinforcing agents, mineral-enriched agent containing essential minerals, more animals and plants extract, seaweed extract, such as physiologically active ingredient of natural ingredients such as herbal components.

As vitamins that can be blended, those listed as fortifiers in food additives can be used, such as vitamin A or β-carotene, vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , vitamin B ₁₂ , Folic acid, pantothenic acid, niacin, biotin, vitamin C, vitamin D, vitamin E, vitamin K, derivatives thereof, oil-coated "vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , vitamin B ₁₂ , folic acid, pantothenic acid, Niacin, biotin, vitamin C ", coenzyme Q ₁₀ (vitamin Q) and the like. Specific examples of derivatives of vitamins, thiamine hydrochloride as vitamin B _1, thiamine sulfate, dibenzoyl thiamine, dibenzoyl thiamine hydrochloride, thiamine nitrate, thiamine laurylsulfate, etc. bisbentiamine, riboflavin as vitamin B ₂ Examples include butyric acid ester and riboflavin 5′-phosphate sodium salt, pyridoxine hydrochloride as vitamin B ₆ , and L-ascorbic acid stearate and sodium L-ascorbate as vitamin C.

  As calcium-containing compounds that can be blended, those listed as calcium fortifiers in food additives can be used, such as calcium chloride, calcium citrate, calcium gluconate, calcium glycerophosphate, calcium acid pyrophosphate, calcium hydroxide, carbonate Calcium, calcium lactate, calcium sulfate, monocalcium phosphate, dicalcium phosphate, tricalcium phosphate, calcium pantothenate, dihydrogen pyrophosphate, coral calcium, dolomite, eggshell calcium, beef bone powder calcium, scallop shell calcium, milk calcium, etc. Can be mentioned.

As iron-containing compounds that can be blended, those listed as iron fortifiers in food additives can be used, such as ferric chloride, iron citrate, ammonium iron citrate, sodium iron citrate succinate, iron lactate Ferrous pyrophosphate, ferric pyrophosphate, ferrous sulfate, ferrous gluconate, heme iron, liver powder, iron coated with fats and oils, and the like.
Moreover, as said essential mineral, zinc, potassium, magnesium, manganese, phosphorus, sodium, selenium, iodine, molybdenum etc. are mentioned, for example. As a source for supplying such essential minerals, for example, wheat extract, wheat germ, young wheat leaf, chlorella, oyster meat extract, seawater concentrate, nuts, fish meal, liver powder, brown rice powder, brewer's yeast, etc. can be used. .

Specific examples of physiologically active ingredients of natural ingredients such as animal and plant extract ingredients, seaweed extract ingredients, herbal medicine ingredients that can be blended are: Ashitaba extract, Avocado extract, Achacha extract, Altea extract, Arnica extract, Aloe extract, Apricot extract, Apricot kernel extract, Ginkgo biloba extract, Fennel extract, Turmeric extract, Oolong tea extract, Ages extract, Echinacea leaf extract, Ogon extract, Prunus extract, Auren extract, Barley extract, Hypericum extract, Odrianthus extract, Dutch mustard extract, Orange extract, Seawater dried , Seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, kawara mugi extract, licorice extract, oil-soluble licorice extract, calcade extract, oyster extract, ki Extract, Kina extract, Cucumber extract, Guanosine, Gardenia extract, Kumazasa extract, Clara extract, Walnut extract, Grapefruit extract, Clematis extract, Chlorella extract, Mulberry extract, Gentian extract, Black tea extract, Yeast extract, Burdock extract, Fermented rice bran extract, Rice germ Oil, Comfrey extract, Collagen, Cowberry extract, Saishin extract, Psycho extract, Saitai extract, Salvia extract, Salmon extract, Sasa extract, Hawthorn extract, Salamander extract, Shiitake extract, Giant extract, Shikon extract, Perilla extract, Linden extract, Shimotake Extract, Peonies Extract, Ginger Root Extract, Birch Extract, Horsetail Extract, Kizuta Extract, Hawthorn Extract, Niwa Coconut extract, Achillea millefolium extract, Pepper mint extract, Sage extract, Zeni mushroom extract, Sensu extract, Senbu extract, Soybean extract, Taisu extract, Thyme extract, Tea extract, Clove extract, Chigaya extract, Chimpi extract, Toki extract, Tokinsenka extract, Tonin extract, Spruce Extract, Dokudami Extract, Tomato Extract, Natto Extract, Carrot Extract, Garlic Extract, Novara Extract, Hibiscus Extract, Bacmond Extract, Parsley Extract, Honey, Hamamelis Extract, Parietalia Extract, Hikikoshi Extract, Bisabolol, Biwa Extract, Fukitane Popo Extract, Fukinoto Extract, Bukuryu Extract, Butcher Bloom Extract, Grape Extract, Placenta Extract, Propolis, Loofah Extract, Beniba Na extract, peppermint extract, body extract, button extract, hop extract, pine extract, horse chestnut extract, mizuba sho extract, mukuroji extract, melissa extract, peach extract, cornflower extract, eucalyptus extract, yukinoshita extract, yuzu extract, yookuin extract, mugwort extract, lavender extract, An apple extract, a lettuce extract, a lemon extract, a forsythia extract, a rose extract, a rosemary extract, a Roman chamomile extract, a royal jelly extract, etc. can be mentioned.

In addition to the above components, sweeteners, coloring agents, seasonings, bittering agents, toughening agents, surfactants, solubilizers, thickeners, pastes, excipients commonly used in foods and pharmaceuticals, etc. , Preservatives, fragrances, antibacterial agents, bactericides, salts, solvents, chelating agents, neutralizing agents, acidulants, pH adjusters, preservatives, buffering agents, fats and oils, processed oils, starch, wheat flour, buckwheat flour, flour Ingredients such as eggs, dairy products, and dairy products can be used.

  Examples of excipients that can be preferably used include agar, gelatin, guar gum, starch, dextrin, cellulose, chocolate, cacao butter, palm oil, palm kernel oil, hardened oil and other fats, various proteins, wheat flour, rice flour , Flour such as red bean flour, soy flour, wheat germ, soy protein, skim milk powder, casein, albumin, globulin, cheese and other dairy products and dairy products, egg white, egg yolk and other egg products, sugar, glucose, maltose , Sugars such as lactose, salts such as sodium chloride and calcium carbonate, etc., and a mixture or composite material thereof may be used.

  Next, examples of cosmetics in which the β-glucan and β-glucan composition of the present invention can be used or formulated include skin lotions, emulsions, skin milks, creams, ointments, external preparations, lotions, calamine lotions, sunscreen agents, suntanes. Basic cosmetics such as preparations, after-shave lotions, pre-shave lotions, makeup bases, packs, cleansings, face wash, anti-acne cosmetics, essences; foundations, white powder, eye shadows, eyeliner, eyebrow, teak, lipstick, nail Makeup cosmetics such as color; shampoo, rinse, conditioner, hair color, hair tonic, set agent, hair conditioner, hair restorer, body powder, deodorant, hair remover, massage cosmetics, body cosmetics, skin cosmetics , Aesthetic cosmetics, pollen Remedies for allergies and allergies, antiallergic agents, rough skin prevention agents, skin disease remedies, skin damage repair agents, wrinkle protection agents, cold compresses, hot compresses, poultices, patches, scrub cosmetics, soaps , Tissue paper, body shampoo, hand soap, perfume, toothpaste, oral care products, mouthwash, gingival massage cream, bath preparation and the like.

  When blended in the above cosmetics, a substance that imparts some physiological activity to the skin may be blended. For example, whitening ingredients, anti-inflammatory agents, anti-aging agents, slimming agents, tanning agents, antioxidants, hair growth agents, hair growth agents, blood circulation promoters, moisturizing ingredients other than polyhydric alcohols or sugars, drying agents, cooling agents , Warming agents, amino acids, wound healing promoters, stimulation relieving agents, analgesics, cell activators, antiallergic agents, rough skin prevention agents, skin damage repair agents, enzyme components, and the like.

  Examples of substances that give physiological activity to the skin include physiologically active ingredients such as animal and plant extract ingredients, seaweed extract ingredients and herbal medicine ingredients, such as Ashitaba extract, Avocado extract, Achacha extract, Altea extract, Arnica extract, Aloe Extract, apricot extract, apricot kernel extract, ginkgo biloba extract, fennel extract, turmeric extract, oolong tea extract, AGES extract, Echinacea leaf extract, Ogon extract, Duckweed extract, Ouren extract, barley extract, Hypericum extract, Odrianthus extract, Dutch mustard extract, Orange extract, seawater dried product, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, cormorant mugwort extract, licorice extract, oil-soluble licorice extract, calcade extract, Kyoku Extract, Kiwi Extract, Kina Extract, Cucumber Extract, Guanosine, Gardenia Extract, Kumazasa Extract, Clara Extract, Walnut Extract, Grapefruit Extract, Clematis Extract, Chlorella Extract, Mulberry Extract, Gentianana Extract, Tea Extract, Yeast Extract, Burdock Extract, Rice Bran Ferment Extract, Rice germ oil, Comfrey extract, Collagen, Cowberry extract, Saishin extract, Psycho extract, Saitai extract, Salvia extract, Sabon extract, Sasa extract, Hawthorn extract, Salamander extract, Shiitake extract, Giant extract, Shikon extract, Perilla extract, Linden extract, Citrus extract, Peonies extract, Ginger root extract, Birch extract, Horsetail extract, Kizuta extract, Hawthorn , Elderberry extract, yarrow extract, mint extract, sage extract, mallow extract, nematode extract, assembly extract, soybean extract, taisu extract, thyme extract, tea extract, clove extract, chigaya extract, chimpi extract, toki extract, tokisenka extract , Tonin extract, Spruce extract, Dokudami extract, Tomato extract, Natto extract, Carrot extract, Garlic extract, Novara extract, Hibiscus extract, Bacmond extract, Parsley extract, Honey, Hamamelis extract, Parietalia extract, Hikikoshi extract, Bisabolol, Biwa extract, Japanese dandelion extract, Japanese quince extract, Bukuryo extract, Butcher bloom extract, Grape extract, Placenta extract, Propolis, F Chima extract, safflower extract, peppermint extract, body extract, button extract, hop extract, pine extract, maronier extract, honeysuckle extract, mukuroji extract, melissa extract, peach extract, cornflower extract, eucalyptus extract, yukinoshita extract, yuzu extract, yokoinin extract, mugwort extract, A lavender extract, an apple extract, a lettuce extract, a lemon extract, a forsythia extract, a rose extract, a rosemary extract, a Roman chamomile extract, a royal jelly extract and the like can be mentioned.

Specific examples of preferable physiologically active components such as other naturally derived components include deoxyribonucleic acid, raffinose, mucopolysaccharide, hyaluronic acid or a salt thereof (such as sodium hyaluronate), sodium chondroitin sulfate, collagen, elastin, chitin, Biopolymers such as chitosan and hydrolyzed eggshell membranes; amino acid derivatives such as amino acids, sarcosine, N-methyl-L-serine; polyhydric alcohols such as ethyl glucose, sodium lactate, urea, sodium pyrrolidonecarboxylate, betaine, whey, or Moisturizing ingredients other than sugars; oily ingredients such as sphingolipids, ceramides, cholesterol, cholesterol derivatives, phospholipids; ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, guaiazulene, hydrocortisone Vitamins such as vitamin A, B ₂ , B ₆ , D, calcium pantothenate, biotin, nicotinamide, vitamin E; allantoin, diisopropylamine dichloroacetic acid, 4-aminomethylcyclohexanecarboxylic acid, etc. Active ingredients: Antioxidants such as carotenoids, flavonoids, tannins, lignans, saponins; cell activators such as α-hydroxy acids, β-hydroxy acids, mevalonic acids, blood circulation promoters such as γ-oryzanol, retinol, retinol derivatives, etc. Whitening agents such as ascorbic acids, arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, glutathione; cephalanthin, licorice extract, red pepper tincture, hinokitiol, garlic iodide, hydrochloric acid Pi Doxin, nicotinic acid, nicotinic acid derivative, calcium pantothenate, D-pantothenyl alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropylmethylphenol, estradiol, ethinylesteradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, Takanal, camphor, salicylic acid, nonyl acid vanillylamide, nonanoic acid vanillylamide, piroctone olamine, pentadecanoic acid glyceryl, l-menthol, mononitroguaiacol, resorcin, γ-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormone, can Talistin tincture, cyclosporine, zinc pyrithione, hydrocortisone, minoxidil, polyoxyethyl monostearate Examples include hair restorers such as sorbitol, mint oil, and Sasanishiki extract, and herbs such as fucoidan and echinesia.

  Examples of the ascorbic acids include ascorbic acid, ascorbic acid sulfate, ascorbic acid phosphate, ascorbic acid higher fatty acid ester, and salts thereof. These salts are sodium salt, potassium salt, magnesium salt, calcium salt, barium salt, ammonium salt, monoethanolamine salt, diethanolamine salt, triethanolamine salt, monoisopropanolamine salt, diisopropanolamine salt, triisopropanolamine. Such as salt. Examples of the ascorbic acid sulfate include ascorbic acid-2-sulfate and ascorbic acid-3-sulfate, and examples of the ascorbic acid phosphate include ascorbic acid-2-phosphate and ascorbic acid. -3-phosphate esters are known, and these are known substances, and are described in JP-B-44-31237 and JP-B-54-21415. Examples of the higher fatty acid esters of ascorbic acid include ascorbic acid-2-palmitic acid monoester, ascorbic acid-2,6-palmitic acid diester, and ascorbic acid-2-stearic acid ester.

  In addition to the above components, oils, powders (pigments, dyes, resins), fluorine compounds, resins, surfactants, stickers, preservatives, fragrances, antibacterial agents, and bactericides that are commonly used in cosmetics Ingredients such as salts, solvents, chelating agents, neutralizing agents, pH adjusting agents and insect repellents can be used.

  Examples of the powder include pigments such as Red No. 104, Red No. 201, Yellow No. 4, Blue No. 1, and Black No. 401, Lake No. 4 such as Yellow No. 4 Al lake, Yellow No. 203 Ba lake, nylon powder, Polymers such as silk powder, urethane powder, Teflon (registered trademark), silicone powder, polymethyl methacrylate powder, cellulose powder, silicone elastomer spherical powder, polyethylene powder; yellow iron oxide, red iron oxide, black iron oxide, Colored pigments such as chromium oxide, carbon black, ultramarine, and bitumen; white pigments such as zinc oxide, titanium oxide, and cerium oxide; extender pigments such as talc, mica, sericite, kaolin, and plate-like barium sulfate; pearls such as mica titanium Pigment: Barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate , Metal salts such as magnesium silicate; inorganic powder such as silica, alumina; bentonite, smectite, and boron nitride. The shapes of these powders include a spherical shape, a rod shape, a needle shape, a plate shape, an indefinite shape, a flake shape, and a spindle shape.

These powders are conventionally known surface treatments such as fluorine compound treatment, silicone treatment, silicone resin treatment, pendant treatment, silane coupling agent treatment, titanium coupling agent treatment, oil agent treatment, N-acylated lysine treatment, The surface treatment may or may not be performed in advance by polyacrylic acid treatment, metal soap treatment, amino acid treatment, inorganic compound treatment, plasma treatment, mechanochemical treatment, or the like.
Among these powders, silicone elastomer spherical powder, polyethylene powder, polypropylene powder, Teflon (registered trademark) powder, silicone rubber, urethane powder, polyalkylsilsesquioxane, nylon, silica beads, alumina beads, apatite, allyl Spherical powders (including hollow resin powders) such as fluorinated acrylic beads are preferably blended because they retain the physiologically active component and are excellent in sustained release effect.

  Examples of the oil agent include volatile and non-volatile oil agents, solvents, and resins that are usually used in skin cosmetics. The oil agent may be a liquid, a paste, or a solid at room temperature, but a liquid that is excellent in handling is preferable. Examples of volatile and non-volatile oils include, for example, higher alcohols such as cetyl alcohol, isostearyl alcohol, lauryl alcohol, hexadecyl alcohol, octyldodecanol; fatty acids such as isostearic acid, undecylenic acid, oleic acid; glycerin, Polyhydric alcohols and sugars such as sorbitol, ethylene glycol, propylene glycol, polyethylene glycol; myristyl myristate, hexyl laurate, decyl oleate, isopropyl myristate, hexyl decyl dimethyloctanoate, glyceryl monostearate, diethyl phthalate, mono Esters such as ethylene glycol stearate and octyl oxystearate; hydrocarbons such as liquid paraffin, petrolatum, squalane; lanolin, Lanolin, waxes such as carnauba wax; and an ethylene-alpha-olefin co-oligomer; mink oil, cacao butter, coconut oil, palm kernel oil, camellia oil, sesame oil, castor oil, oils such as olive oil.

  Examples of other forms of oil include, for example, dimethylpolysiloxane, methylhydrogenpolysiloxane, methylphenylpolysiloxane, polyether-modified organopolysiloxane, fluoroalkyl / polyoxyalkylene co-modified organopolysiloxane, alkyl-modified Silicone compounds such as organopolysiloxane, terminal-modified organopolysiloxane, fluorine-modified organopolysiloxane, amodimethicone, amino-modified organopolysiloxane, silicone gel, acrylic silicone, trimethylsiloxysilicate, silicone RTV rubber; perfluoropolyether, fluorinated Fluorine compounds such as pitch, fluorocarbon, and fluoroalcohol are listed.

  Examples of the solvent include purified water, cyclic silicone, ethanol, light liquid isoparaffin, lower alcohol, ethers, LPG, fluorocarbon, N-methylpyrrolidone, fluoroalcohol, volatile linear silicone, and next-generation fluorocarbon. It is done.

  As the surfactant, for example, an anionic surfactant, a cationic surfactant, a nonionic surfactant, or a betaine surfactant can be used.

  Examples of the above-mentioned adhesives and resins include sodium polyacrylate, cellulose ether, calcium alginate, carboxyvinyl polymer, ethylene / acrylic acid copolymer, vinyl pyrrolidone polymer, vinyl alcohol / vinyl pyrrolidone copolymer, nitrogen substitution Acrylamide polymers, polyacrylamides, cationic polymers such as cationized gar gum, dimethylacrylammonium polymers, acrylic methacrylic acid acrylic copolymers, POE / POP copolymers, polyvinyl alcohol, pullulan, agar, gelatin, tamarind seeds Sugar, xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, gar gum, gum arabic, crystalline cellulose, arabinogalactan, karaya gum, tragacanth , Alginic acid, albumin, casein, curdlan, gellan gum, dextran, cellulose, polyethyleneimine, highly polymerized polyethylene glycol, cationized silicone polymer, synthetic latex, and the like.

It is also preferable to impart a UV protection effect to cosmetics. In this case, it is preferable to blend an ultraviolet protective agent (also referred to as an ultraviolet absorber) as shown below.
Examples of ultraviolet protective agents (including organic and inorganic systems, which may correspond to either UV-A or B) include fine particle titanium oxide and fine particle zinc oxide. Examples of the organic ultraviolet protective agent include 2-methoxyhexyl paramethoxycinnamate, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfate, and 2,2′-dihydroxy-4- Methoxybenzophenone, p-methoxyhydrocinnamic acid diethanolamine salt, paraaminobenzoic acid (hereinafter abbreviated as PABA), ethyldihydroxypropyl PABA, glyceryl PABA, homomenthyl salicylate, methyl-O-aminobenzoate, 2-ethylhexyl-2-cyano- 3,3-diphenyl acrylate, octyl dimethyl PABA, octyl methoxycinnamate, octyl salicylate, 2-phenyl-benzimidazole-5-sulfate, triethanolamine salicylate, 3- (4-methylbenzili Den) camphor, 2,4-dihydroxybenzophenine, 2,2 ', 4,4'-tetrahydroxybenzophenone, 2,2'-dihydroxy-4,4'-dimethoxybenzophenone, 2-hydroxy-4-N- Octoxybenzophenone, 4-isopropyldibenzoylmethane, butylmethoxydibenzoylmethane, 4-tert-butyl-4'-methoxydibenzoylmethane, 4- (3,4-dimethoxyphenylmethylene) -2,5-dioxo-1 -Imidazolidinepropionate 2-ethylhexyl, polymer derivatives thereof, silane derivatives and the like. Furthermore, what sealed these in the polymer may be used.

  Examples of the β-glucan and the chemical product in which the β-glucan composition of the present invention can be used or blended include medical detergents, clothing detergents, contact lens preservatives, contact lens cleaning liquids, disinfectants, soaps, hygiene products, medical supplies , Toiletries, thickeners, viscosity modifiers and the like.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

Analysis Example 1 (Confirmation of β-glucan and measurement of content)
For analysis of β-glucan, the total amount of polysaccharide precipitated by alcohol is measured by the phenol-sulfuric acid method, and confirmation and quantification of β-glucan in the precipitated polysaccharide is performed by Seikagaku Corporation (1-3) -β. This was carried out using a kit for detecting and measuring β-glucan containing -D-bonding. First, the total polysaccharide amount in the measurement sample was measured by the phenol sulfuric acid method. That is, 30 μl of distilled water is added to 30 μl of the sample solution, 120 μl of a phosphate buffer solution (pH 6.9) containing 300 mM NaCl is added thereto, 640 μl of ethanol (3 times volume) is further added, and the mixture is kept at −15 ° C. for 10 minutes. The polysaccharide was left to stand. After removing the supernatant, 100 μl of distilled water was added and dissolved. To this, 100 μl of 5 wt% aqueous phenol solution and 500 μl of sulfuric acid were added and reacted. Absorbance at 490 nm was measured using a blank obtained by adding phenol solution and sulfuric acid to 100 μl of distilled water without adding a sample. A calibration curve was prepared using a 2-fold dilution series prepared from pullulan 10 mg / ml as a standard sample, and the amount of polysaccharide was quantified. Next, a solution having a total polysaccharide amount of about 1 to 0.1 mg / ml is first diluted 10-fold with 0.5 M NaOH, and then diluted with β-glucan-free distilled water, and diluted to 10 ⁻¹⁰ Was prepared. 50 μl of β-glucan diluted solution was taken in a tube, 50 μl of the main reaction reagent was added, and incubated at 37 ° C. for 30 minutes. Subsequently, 50 μl of sodium nitrite solution, 50 μl of ammonium sulfamate, and 50 μl of N methyl 2-pyrrolidone solution were added and reacted, and the absorbance of the solution was measured at 545 nm (target wavelength: 630 nm). A calibration curve was obtained with a β-glucan solution of 7.5-60 pg / ml with the attached β-glucan standard, and the concentration of each β-glucan solution was calculated.

Analysis Example 2 (Measurement of molecular weight)
The molecular weight of β-glucan was measured as follows. That is, 3 times the amount of alcohol was added to the β-glucan solution, cooled to −20 ° C. and allowed to stand for 10 minutes to obtain a precipitate. 5 mg of the precipitated β-glucan precipitate was taken in a tube, dissolved by adding 1 ml of distilled water, and dissolved by adding 3 times the amount of ethanol. A solution obtained by repeating this operation three times was dialyzed against water and freeze-dried to obtain purified β-glucan. β-glucan was dissolved in distilled water, and the concentration was measured by the phenol sulfuric acid method. The calibration curve of the phenol sulfuric acid method was pullulan. For separation, Shodex packed columns KS-804 and KS-802 (manufactured by Showa Denko), which are HPLC gel filtration columns, were used at a flow rate of 0.5 ml / min. The temperature was 80 ° C., and a molecular weight measuring device TriSEC Model 302 manufactured by Viscotech was used for detection, and three points of differential pressure viscosity, differential refraction, and light scattering were measured, and the molecular weight was calculated from the three pieces of information. First, a solution (1.5 mg / ml, 2.5 mg / ml, 5.0 mg / ml) with an accurate β-glucan concentration was prepared, and the refractive index specific to the sample was calculated from the sample concentration and the differential refractive area value. did. In the measurement of the differential pressure viscosity, pullulan was used as a standard material for the intrinsic viscosity. The approximate molecular weight is calculated from the differential refraction detection area value and the light scattering detection area value measured for the purified β-glucan solution (1.5 mg / ml), and the molecular weight is corrected from the calculated intrinsic viscosity of β-glucan. Of molecular weight.

Analysis example 3 (analysis of constituent sugars)
Analysis of the constituent sugars of β-glucan was performed by the alditol acetate method. The polysaccharide purified by alcohol precipitation was dissolved in water and dialyzed to completely remove monosaccharides / oligosaccharides. The presence or absence of a low molecular sugar was confirmed by HPLC. In acid hydrolysis of purified β-glucan, 0.1 ml of 72 wt% sulfuric acid was added to 1 mg of purified β-glucan, and the mixture was heated at 100 ° C. for 4 hours after ice-cooled reaction. The mixture was neutralized and centrifuged to collect the supernatant, which was desalted and concentrated with a vacuum evaporator. Next, alditolization was performed. That is, 1 ml of distilled water was added to the concentrated sample and 1 drop of 2N ammonia water was added. Then, 3 mg of sodium borohydride was added and reacted at room temperature for 2 hours. The mixture was neutralized with an ion exchange resin and concentrated with a vacuum evaporator. Further, 1 ml of methanol was added and concentrated with a vacuum evaporator. Next, 0.1 ml of acetic anhydride and 0.1 ml of pyridine were added to the concentrated sample and reacted at 100 ° C. for 2 hours. 1 ml of distilled water was added and the mixture was concentrated with a vacuum evaporator. This concentration operation was repeated three times, 20 μl of acetone was added to the dried sample, and measurement was performed by gas chromatography. For gas chromatography, a capillary column TC-WAX (GL Sciences) is used and a carrier gas He is used.
110 kPa, a column temperature of 230 ° C., an inlet temperature of 250 ° C., and a detection method was hydrogen ionization detection, and 5890A Gas Chromatography (manufactured by HEWLETT PACKARD) was used. The constituent sugar composition was calculated from the peak area ratio of the measurement data. In this analysis, inositol was used as an internal standard, and it was confirmed that acetylated derivatives of monosaccharides could be completely separated and quantified in a mixture of glucose, mannose, galactose, rhamnose, fucose, xylose, arabinose, and ribose.

Analysis Example 4 (Analysis of binding position)
Analysis of the binding mode of β-glucan was performed by the methylation method. DMSO (2 ml) was added to and dissolved in 3 mg of β-glucan purified in the same manner as in Analytical Example 3, 0.5 ml of carbanion reagent (adjusted with DMSO and sodium hydride) was added, and the mixture was reacted at room temperature for 4 hours. While cooling, 1.5 ml of methyl iodide was added and allowed to react for 1.5 hours. The reaction solution was dialyzed against running water for 3 hours and overnight with 200-fold distilled water, and then concentrated to dryness with a vacuum evaporator. After this methylation operation was repeated three times, acid hydrolysis and alditol acetate formation were performed in the same manner as in Analysis Example 3. To the obtained sample, 20 μl of acetone was added and detected by gas chromatography. Each sugar derivative was quantified from the peak area ratio of the measurement data. In this analysis, inositol was used as an internal standard, 2,3,4,6 methyl 1 acetyl glucose, 2,4,6 methyl 1,3 acetyl glucose, 2,3,6 methyl 1,4 acetyl glucose, 2, It was confirmed that each derivative could be completely separated and quantified in a mixture of 3,4 methyl 1,6 acetyl glucose, 2,4 methyl 1,3,6 acetyl glucose and 2,3 methyl 1,4,6 acetyl glucose.

Analysis Example 5 (Quantitative analysis method of 1,3,1,4-β-D-glucan)
1,3,1,4-β-D-glucan was quantified by the McCleary method (enzyme method) using a β-glucan measurement kit manufactured by Megazyme. First, the water content of the measurement sample was measured, 10 mg of the sample was taken in a 17 ml tube, and 200 μl of a 50% ethanol solution was added and dispersed. Next, 4 ml of 20 mM phosphate buffer (pH 6.5) was added and mixed well, and then heated in a boiling water bath for 1 minute. Mix well and heat in hot water bath for another 2 minutes. After cooling to 50 ° C. and allowing to stand for 5 minutes, add 200 μl (10 U) of the lichenase enzyme solution (diluted with 20 ml of 20 mM phosphate buffer, the remaining amount is stored frozen) to each tube, The reaction was carried out at 50 ° C. for 1 hour. 5 ml of 200 mM acetate buffer (pH 4.0) was added to the tube and mixed gently. It was left to stand at room temperature for 5 minutes, and the supernatant was obtained by centrifugation. Take 100 μl in three tubes, one with 100 μl 50 mM acetate buffer (pH 4.0), the other two with 100 μl (0.2 U) β-glucosidase solution (20 ml of vial attached to kit) Diluted with 50 mM acetic acid buffer solution, and the remaining amount was stored frozen), and reacted at 50 ° C. for 10 minutes. 3 ml of glucose oxidase / beloxidase solution was added and reacted at 50 ° C. for 20 minutes, and the absorbance (EA) at 510 nm of each sample was measured. β-glucan content was determined by the following formula.
β-glucan content (% by weight) = (EA) × (F / W) × 8.46
F = (100) / (absorbance of glucose 100 μg)
W = calculated anhydride weight (mg)

Example 1 (Production of β-glucan)
FERM BP-8391 strain was inoculated into 120 ml of YM medium (manufactured by Difco) to obtain a preculture (26 ° C., 180 rpm, 3 days culture). Sucrose 90 g, yeast extract _{15g, K 2 HPO 4 1.0g,} KCl 0.5g, MgSO 4 · 7H 2 O 0.5g, 5L jar containing the FeSO ₄ · 7H ₂ O 0.01g and distilled water 3L A mentor was inoculated with 100 ml of the preculture solution and cultured at 26 ° C. for 72 hours to obtain 3 L (pH 4.5) of the culture solution. The culture solution was sterilized by heating at 80 ° C. for 30 minutes, added with an equal amount of distilled water, mixed well, and then centrifuged at 8000 rpm for 30 minutes to obtain a culture supernatant (A). An equal volume of ethanol was added to 200 ml of the culture supernatant (A), and the resulting polysaccharide precipitate was separated, washed with ethanol, and dissolved in 100 ml of distilled water. After removing insoluble matter, 200 ml of ethanol was added to obtain a precipitate. This operation was repeated again, and the polysaccharide solution dissolved in water was put into a dialysis membrane (molecular weight 3000 cut), dialyzed with 100 times the amount of distilled water, and freeze-dried to obtain 16 g of culture supernatant powder (B).

Example 2 (Production of β-glucan)
The FERM BP-8391 strain was inoculated into 120 ml of a medium containing 1% yeast extract (manufactured by Difco) and 1% by weight glucose to obtain a preculture solution (cultured at 26 ° C., 180 rpm, 3 days). Sucrose 90 g, yeast extract _{2g, K 2 HPO 4 1.0g,} KCl 0.5g, MgSO 4 · 7H 2 O 0.5g, 5L jar containing the FeSO ₄ · 7H ₂ O 0.01g and distilled water 3L A mentor was inoculated with 100 ml of the preculture solution and cultured at 26 ° C. for 72 hours to obtain 3 L (pH 4.5) of the culture solution. The culture solution was sterilized by heating at 80 ° C. for 30 minutes, an equal amount of distilled water was added and mixed well, and then centrifuged at 8000 rpm for 30 minutes to obtain a culture supernatant (C). An equal amount of ethanol was added to 200 ml of the culture supernatant (C), and the resulting polysaccharide precipitate was separated, washed with ethanol, and dissolved in 100 ml of distilled water. After removing insoluble matter, 200 ml of ethanol was added to obtain a precipitate. This operation was repeated again, and the polysaccharide solution dissolved in water was placed in a dialysis membrane (molecular weight 3000 cut), dialyzed with 100 times the amount of distilled water, and freeze-dried to obtain 10 g of culture supernatant powder (D).

Example 3 (Production of β-glucan)
Tunicase (manufactured by Daiwa Kasei Co., Ltd.) was added to the culture broth obtained in Example 1 so as to be 0.05% by weight, stirred at 40 ° C. for 2 hours, and then sterilized by heating at 80 ° C. for 30 minutes. . Two times the amount of distilled water was added thereto, mixed well, and then centrifuged at 8000 rpm for 30 minutes to obtain a culture supernatant (E). An equal amount of ethanol was added to 300 ml of the culture supernatant (E), and the resulting polysaccharide precipitate was separated, washed with ethanol, and dissolved in 100 ml of distilled water. After removing insoluble matter, 200 ml of ethanol was added to obtain a precipitate. This operation was repeated again, and the polysaccharide solution dissolved in water was put into a dialysis membrane (molecular weight 3000 cut), dialyzed with 100 times amount of distilled water, and freeze-dried to obtain 18 g of culture supernatant powder (F).

Example 4 [Production of β-glucan composition (β-glucan complex) with 1,3,1,4-β-D glucan]
10 ml of distilled water was added to 100 mg of the culture supernatant powder (B) obtained in Example 1 and dissolved well. 10 ml of distilled water was added to 100 mg of 1,3,1,4-β-D glucan β-glucan MW standard (molecular weight 40,000, manufactured by Megazyme) and dissolved well. Both prepared solutions were mixed well and then lyophilized to obtain β-glucan composition powders (G), (H) and (I), respectively. The mixing ratio of the culture supernatant powder (B) and the β-glucan MW standard was as follows by weight ratio. 4: 1 (G), 1: 1 (H), 1: 4 (I).

Comparative Example 1
The same procedure as in Example 1 was carried out except that the IFO4466 strain was used instead of the FERM BP-8391 strain, to obtain 1.5 g of a culture supernatant (J) and a culture supernatant powder (K).

Test Example 1 (Solubility test)
Samples obtained in Examples [culture supernatant powders (B), (D) and (F), β-glucan composition powders (G), (H) and (I)] and samples obtained in Comparative Examples A solubility test was performed on [culture supernatant powder (K)]. In addition, as a control, schizophyllan (pharmaceutical name Sonifilan: manufactured by Kaken Pharmaceutical Co., Ltd.) and curdlan (manufactured by Wako Pure Chemical Industries) were evaluated together. 10 mg of each powder sample was taken in a 1.5 ml tube and 1 ml of distilled water was added. The tube was set on a rotator RT-50 (manufactured by Taitec Co., Ltd.), which is a rotary stirrer, and the degree of dissolution was observed with the naked eye while rotating at 30 rpm at room temperature. Observations were made after 5 minutes, 10 minutes, 30 minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes, 360 minutes, 720 minutes, and 1440 minutes, and the results are shown in Table 1. It was.
Evaluation is that the solution is uniformly and completely dissolved (◎), hydrated and swollen into a transparent liquid, but non-uniform parts are observed (○), parts that are not dissolved (△), It was classified into four points, that is, a precipitate was observed and it was not dissolved (×).

Test example 2 (molecular weight measurement)
The molecular weight of the samples [culture supernatant powders (B), (D) and (F)] obtained in Examples and the sample [culture supernatant powder (K)] obtained in Comparative Examples was measured according to Analysis Example 2. The results are shown in Table 2. As a result, schizophyllan has a molecular weight of 440,000 and a value almost identical to the literature value, and β-glucan [culture supernatant powders (B), (D) and (F)] obtained from FERM BP-8391 strain, Both were measured to have a molecular weight of less than 500,000.

Test Example 3 (Structural sugar analysis results)
Constituent sugar analysis was performed according to Analysis Example 3, and the results are summarized in Table 3. FERM
Β-glucan [culture supernatant powders (B), (D) and (F)] obtained from the BP-8391 strain is β-glucan with glucose of 80% or more, and culture supernatant powders (B) and (F) Was confirmed to be a polysaccharide containing mannose.

Test example 4 (analysis result of bonding position)
According to Analysis Example 4, the binding mode was analyzed, and the peak area value and the ratio of glucose residues having only 1,3 bonds to the total glucose residues involved in binding between glucose are shown in Table 4. It was confirmed that β-glucan [culture supernatant powders (B), (D) and (F)] obtained from the FERM BP-8391 strain is a highly branched polysaccharide.

Test Example 5 (Quantification of 1,3,1,4-β-D-glucan)
For the samples [β-glucan composition powders (G), (H) and (I)] obtained in Examples, 1,3,1,4-β-D-glucan was quantified. When measured according to Analysis Example 5, the amount of 1,3,1,4-β-D-glucan in the sample was calculated as (G) 20%, (H) 46%, (I) 80%, and the theoretical value. Values approximating (25%, 50%, 75%) were obtained.

Test Example 6 (Evaluation of immune enhancing activity by injection administration to mice)
Using the β-glucan culture supernatant powder (B) obtained in Example 1, immunostimulatory activity was measured. 0.04 mg, 0.2 mg, 1 mg and 5 mg of the culture supernatant powder (B) were dissolved in 0.25 ml of physiological saline and administered intraperitoneally to ICR mice (female, 4 weeks old). 6 hours after administration, peritoneal cells were collected, centrifuged, and stained to determine the number of intraperitoneal cells and the number of neutrophils. The measurement results are shown in FIG. 1 and Table 5, assuming that the number of intraperitoneal cells and the number of neutrophils when the culture supernatant powder (B) was administered at 0.04 mg was 1.

Next, 1 mg of the culture supernatant powder (B) was dissolved in 0.25 ml of physiological saline, and intraperitoneally administered to ICR mice (female, 4 weeks old). The change in the number of balls over time was observed. For comparison, lentinan was used (1 mg administration). The number of intraperitoneal cells and the number of neutrophils after 48 hours was 1, and the measurement results are shown in FIGS.
From the above, it has been clarified that the β-glucan of the present invention exhibits an immune enhancing effect by activating leukocytes.

Test Example 7 (Evaluation of immune enhancement activity by injection administration to mice)
1 mg of each sample [culture supernatant powders (B), (D) and (F), β-glucan composition powder (G), schizophyllan] was dissolved in physiological saline, and ICR mice (female, 4 weeks old) ) Was administered intraperitoneally. 6 hours after administration, peritoneal cells were collected, and the number of intraperitoneal cells and the number of neutrophils were measured. The values when each sample was administered were compared with the number of intraperitoneal cells and the number of neutrophils when sizzophyllan was administered as 1, and the results are shown in Table 7. All of the β-glucans of the present invention tested showed an increase in neutrophils and were found to have the basic property of immune enhancement (BRM activity).

Test Example 8 (Evaluation of immune enhancing activity by oral administration to mice)
The culture supernatant (A) and the culture supernatant powder (B) obtained in Example 1 were each diluted with distilled water to prepare a β-glucan concentration of 2.5 mg / ml. After sterilizing the solution, Ad libitum as drinking water. As a control, the distilled water used for dilution of the sample was used as drinking water. After feeding on a solid feed for 1 week, peritoneal cells were collected, TNF-α was measured after 2 hours of culture, and IL-12 was measured after 24 hours of culture by ELISA. As a result, the oral administration group of the culture supernatant liquid (A) and the culture supernatant powder (B) was 3 times or more in TNF-α and IL-12 was 2 times or more compared to the control group to which only distilled water was administered. Production was observed. From the above, it has been clarified that the β-glucan of the present invention is a β-glucan that exhibits good solubility and excellent functionality for enhancing immunity.

10 is a graph showing the measurement results of the number of intraperitoneal cells and the number of neutrophils 6 hours after administration to mice in Test Example 6. 10 is a graph showing the change over time in the number of intraperitoneal cells after administration to mice in Test Example 6. 10 is a graph showing the change over time in the number of neutrophils after administration to mice in Test Example 6.

Claims (5)

  1,3,1,6-β-D-glucan having a weight average molecular weight of 5000 or more and less than 500,000, and the proportion of β-1,3 bonded glucose residues having no branching is Β-glucan, which is 1% or more and 40% or less.   The β-glucan according to claim 1, wherein the constituent sugar of β-glucan is glucose alone or glucose and mannose.   The β-glucan according to claim 1 or 2, wherein the β-glucan is obtained by culturing Aureobasidium pulullans ADK-34 (FERM BP-8391) strain.   A β-glucan composition comprising the β-glucan according to claim 1 in an amount of 1% by weight or more based on the total amount of β-glucan. The β-glucan composition according to claim 4, comprising the β-glucan according to any one of claims 1 to 3, and a β-glucan derived from a gramineous plant or 1,3,1,4-β-D-glucan.

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