JP2005307150A - Beta glucan - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a β-glucan and a β-glucan complex having excellent physiological regulation function and excellent physiological activation function and provide a method for producing an oligomerized β-glucan that is safe when it is used as a food, a medicine or a cosmetic, very easy to handle since the problems on thickening, or the like, can be controlled when it is produced or used, and in addition, its formulation quantity is not substantially limited because it can be readily powdered or concentrated. <P>SOLUTION: This invention provides β-glucans respectively originating from microorganisms, cereals, Basidiomycetes, and further a glucan complex containing two or more kinds of these β-glucans. In addition, the invention provide a method for producing the oligomerized β-glucan by allowing a hydrolytic enzyme to act on the β-glucans originating from microorganisms, cereals or Basidiomycetes. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to β-glucan and a method for producing β-glucan. The present invention also relates to a water-soluble β-glucan derived from cereal.

  In recent years, β-glucan has an excellent bioregulatory function and bioactive function, for example, lipid metabolism improving action, intestinal regulating action, blood sugar level rise inhibiting action, cholesterol lowering action, blood sugar level lowering action, antitumor action, immune enhancing action, It is a material that has been attracted attention for its analysis of immunostimulatory action, intestinal immunity enhancing action, skin immunity enhancing action, lifestyle-related disease prevention / amelioration action, and the like.

  β-glucan is contained in many organisms, for example, microorganisms, basidiomycetes, cereals and other plants, and is mainly present as a component constituting these cell walls (some microorganisms have β Some also secrete glucans). Its structure is as follows: 1-2β-D-glucopyranose bond, 1-3β-D-glucopyranose bond, 1-4β-D-glucopyranose bond, 1-6-β-D-glucopyranose The main component is a glucose polymer having at least two types of bonds.

  Conventionally, when β-glucan is used, it is only for one type of β-glucan, and a synergistic effect by using two or more types of β-glucan has not been considered at all.

  In addition, β-glucan contained in basidiomycete (mushroom) fruit bodies and mycelium has high immunopotentiating activity, and some of them are used as pharmaceuticals, such as lentinan extracted from shiitake fruit bodies. However, β-glucan contained in the fruiting bodies and mycelium of basidiomycetes (mushrooms) generally varies greatly depending on the growth and cultivation conditions, and it is necessary to separate β-glucan by extraction operation. As a result, β-glucan obtained as a result covers a wide variety of molecular species, and the quality is unstable, such as a mixture of high and low molecular weight substances. Thus, it is difficult for basidiomycetes (mushrooms) to stably produce a certain β-glucan having high activity.

  Moreover, it is well known that cell wall components of microbial cells generally contain β-glucan and have an activity of enhancing immune activity (Patent Documents 1 to 5), which is interesting as a source of β-glucan. However, since the cell wall of microbial cells contains components other than β-glucan and is insoluble in water, extraction and purification operations are necessary to obtain highly effective water-soluble and high-purity β-glucan. . For this purpose, it is necessary to perform an extraction operation after homogenizing microbial cells to self-digest or decomposing the cell wall by adding an enzyme or the like. Since it contains components other than intracellular components derived from lysis and β-glucan derived from cell walls, separation and purification operations become complicated, and establishment of a method for stably extracting β-glucan of a certain quality can be said to be an issue. .

  On the other hand, microorganisms that secrete and produce β-glucan having a function of enhancing immune activity outside the bacterial body are known. Examples include Macrophhomopsis genus (Patent Document 6), Aureobasidium pullulans (Non-patent Document 2, Patent Document 7), or curdlan produced by Alkaligenes (Non-patent Document 1). Can be mentioned. If β-glucan can be fermented and produced using such microorganisms, it is possible to obtain β-glucan that is uniform, highly water-soluble, and has strong immune activity enhancing activity, and is an extremely effective method. Therefore, a method for producing β-glucan using a microorganism belonging to the genus Aureobasidium, which is known to secrete and produce highly active β-glucan outside the cell, has been proposed. (Patent Documents 11, 12, and 13)

  Β-glucan produced by the genus Aureobasidium has both high molecular weight and high branching density. It is known that β-glucan exhibiting antitumor activity has a branched structure, and it is suggested that a higher-order structure derived from molecular weight, branch density, branched chain length, etc. is related (Non-patent Document 3). , P. 7), it can be said that β-glucan is very promising in terms of function. And it is presumed that the branch density is high. For example, it is much more water-soluble than curdlan composed only of β-1,3 bonds, and it can be said that it is a preferable property in terms of physical properties. On the other hand, it is presumed that both the molecular weight and the branch density are high, but the viscosity is very high and thixotropic properties are exhibited. From the user's point of view, this property is difficult to use due to poor handling when used as a raw material or material for foods, pharmaceuticals, cosmetics, etc. It was very problematic in terms of efficiency.

  Therefore, a technique for reducing the molecular weight to an appropriate molecular weight while maintaining the function of β-glucan is expected. β-glucan can be reduced in molecular weight by hydrolysis with acid or alkali, heating method, ultrasonic treatment method, high shear treatment method, high-speed jetting method, gamma irradiation method, oxidation Methods of reducing have been proposed (Patent Documents 8 to 10, Non-Patent Documents 3, 11 pages, Non-Patent Documents 4, 5). Β-glucan produced by Aureobasidium is difficult to be hydrolyzed by physical treatment such as heating, ultrasonic treatment, high shear force treatment, high-speed jet treatment, etc., and gamma-ray irradiation treatment and high-speed jet treatment require large equipment. Since time is required, it is not always an excellent method for supplying inexpensive raw materials and raw materials particularly required for food applications. On the other hand, the treatment with acid or alkali is preferable because the selectivity of the degradation site is low and the production of monosaccharides is remarkable, and when the fermentation broth is used as it is, coloring due to coexisting proteins and the like occurs. Absent. Further, since chemicals are used for the oxidation-reduction treatment, it is not preferable for use as a raw material for foods, pharmaceuticals, and cosmetics.

  On the other hand, although a method using an enzyme such as glucanase or cellulase can be considered, β-glucan produced by aureobasidium is hardly affected by commercially available enzymes for the food industry, and there are no reports. (This is presumably because the action of the enzyme is hindered by steric hindrance because of its high branching density.) Therefore, examples of methods that have been carried out with enzymes so far include alkaline solutions, organic solvents, An example (Patent Document 14) in which an enzyme is allowed to act under conditions using an auxiliary agent such as urea is only recognized. Moreover, although patent document 15 has the description which carries out the enzyme process of the beta-glucan derived from basidiomycetes, the specific example is not disclosed. Patent Document 16 discloses an example in which hemicellulase or hemicellulase and pectinase are used together when β-glucan is extracted from mycelium and fruiting bodies of Agaricus. This is a method for obtaining necessary β-glucan by removing necessary components with an enzyme, which is completely different from the method for hydrolyzing β-glucan of the present invention.

  Β-glucan is a high-molecular polysaccharide in which D-glucose is polymerized by β-bonding, and is a main component of an antitumor polysaccharide contained in mushrooms, and β-glucan contained in cereals has an excellent cholesterol-reducing effect. It is known that In recent years, cereal-derived β-glucan molecular weight-reduced products (extracted with molecular weights reduced to 100,000 or less) have also been recognized as an immunopotentiating effect (Patent Document 17) and added to various foods. In this case, an applied technology has been proposed since it exhibits an excellent effect of improving physical properties (Patent Documents 18 to 22).

  This β-glucan is relatively present in barley and oats, and its content is 6% (Non-patent Document 6). In barley and oats, β-glucan exists as a cell wall component of starch storage cells, and its molecular weight is said to be several million. Β-glucan obtained by extraction by adding a solvent such as water to barley or oats has a lower molecular weight than that present in the grain due to enzymes and physical action originally present in the grain, and its molecular weight is Several hundred thousand to several tens of thousands (Patent Documents 23 and 24).

  These β-glucans have excellent health functionality (physiological functionality) as described above, but have a difficulty in solubility in water. Specifically, the extracted β-glucan is easily soluble in hot water but hardly soluble in cold water. Further, an aqueous solution in which β-glucan is dissolved also has a property that β-glucan is precipitated by freezing and thawing. When the concentration is relatively high, an aqueous solution in which β-glucan is dissolved may be gelled or precipitated by refrigeration (cooling to 10 ° C. or lower).

  On the other hand, the health functionality of β-glucan is due to the excellent water solubility of β-glucan. Therefore, in order to eat β-glucan and cause the β-glucan to exhibit functionality in vivo, it is necessary that β-glucan is dissolved or easily dissolved at the time of feeding. As described above, β-glucan derived from cereal may precipitate and agglomerate depending on the storage and processing form, and this insoluble β-glucan is reheated at the time of feeding in order to exert its original functionality. Therefore, it was necessary to improve the solubility or restore the solubility in a hydrated state. In beverages and the like, the resulting β-glucan precipitation contributed to the deterioration of quality. In order to solve such drawbacks, it has been desired to create β-glucan that does not precipitate even after repeated freezing / thawing or retains the property of dissolving in cold water.

JP 54-138115 A Japanese Patent Laid-Open No. 09-103266 Japanese Patent Laid-Open No. 03-229702 JP-A-10-167972 Japanese Patent Publication No. 06-92441 Japanese Unexamined Patent Publication No. 03-2202 JP-A-6-340701 JP-A-63-77901 JP-A-5-262654 Japanese Unexamined Patent Publication No. 55-71701 Japanese Patent No. 3444424 JP-A-7-51080 JP 2004-49013 A JP-A-6-277085 JP-A-63-77901 Japanese Patent No. 3428356 JP 2001-323001 A JP 2002-241784 A Japanese Patent Laid-Open No. 2002-306056 JP 2002-306124 A JP 2002-306064 A JP 2002-306094 A JP-T-2001-501996 JP 2002-105103 A Science of dietary fiber, p108, Asakura Shoten (1997) Agric. Biol. Chem. 47 (6), 1167-1172 (1983); Structure and physiological activity of polysaccharides, Asakura Shoten (1990) Agric. Biol. Chem. 48 (4), 915-912 (1984); Journal of Japanese Society for Agricultural Chemistry; 66 (11), 1633-1640 (1992) The latest food standard ingredient list, national cook training facility association

Accordingly, an object of the present invention is to provide β-glucan and β-glucan complex having an excellent bioregulatory function and physiologically active function, and is also safe for use in the production of foods, pharmaceuticals and cosmetics, and at the time of production. A method for producing low molecular weight β-glucan that is very easy to handle by suppressing problems such as thickening during use, and that is easy to powder and concentrate and whose target compounding amount is not substantially limited. It is to provide.
Further, the object of the present invention is excellent in solubility in water, stable even when stored in an aqueous solution without causing changes over time such as precipitation or turbidity, and does not precipitate even after repeated freezing / thawing. The object is to provide a water-soluble β-glucan having excellent solubility in cold water.

As a result of intensive studies, the present inventors have completed the present invention.
That is, the present invention provides a β-glucan derived from microorganisms.

  The present invention also provides a β-glucan derived from the microorganisms, which is β1-3, 1-6 glucan.

  The present invention also provides cereal-derived β-glucan.

  The present invention also provides the above-mentioned grain-derived β-glucan, which is β1-3, 1-4 glucan.

  The present invention also provides β-glucan derived from basidiomycetes.

  The present invention also provides a β-glucan derived from the basidiomycete, which is β1-3, 1-6 glucan.

  The present invention also provides a β-glucan complex containing β1-3, 1-6 glucan and β1-3, 1-4 glucan.

  The present invention also provides a β-glucan complex containing at least two of β-glucan derived from microorganisms, β-glucan derived from cereals, and β-glucan derived from basidiomycetes.

  In addition, the present invention includes any one or more of β1-3, 1-6 glucan derived from microorganisms, β1-3, 1-4 glucan derived from grains, and β1-3, 1-6 glucan derived from basidiomycetes. A β-glucan complex is provided.

  The present invention also provides a method for producing a low molecular weight β-glucan derived from microorganisms having a step of allowing a hydrolase to act.

  Moreover, this invention provides the manufacturing method of the low molecular-weight beta-glucan derived from a grain which has the process of making a hydrolase act.

  The present invention also provides a method for producing a low molecular weight β-glucan derived from basidiomycetes having a step of allowing a hydrolase to act.

  Further, as a result of intensive studies on the solubility of β-glucan derived from cereal in water, the present inventors have a close relationship with the molecular weight of the cereal-derived β-glucan, and β-glucan having a specific molecular weight It has been found that an excellent β-glucan can be created by controlling the molecular weight of. As a result of further investigations, the present inventors made a limited hydrolysis reaction by causing an enzyme having hydrolysis activity to act on grains such as barley or oats or β-glucan derived from the grains. It was found that by adjusting the molecular weight of glucan, the solubility of β-glucan can be modified to create an excellent β-glucan.

  The present invention has been made based on the above findings, and provides a water-soluble β-glucan obtained by adding an enzyme to cereal and / or cereal-derived β-glucan to reduce the molecular weight.

  In addition, the present invention provides the water-soluble β-glucan obtained by adding an enzyme to a grain containing hemicellulose and / or a beta-glucan derived from a grain containing hemicellulose to act on it. It is.

  In addition, the present invention provides the water-soluble β-glucan obtained by adding an enzyme to barley or oats and / or β-glucan derived from barley or oats to reduce the molecular weight. .

  The present invention also provides the water-soluble β-glucan, wherein the β-glucan derived from cereal is an extract or fraction extracted from the cereal.

  In addition, the present invention provides the water-soluble β-glucan having a weight average molecular weight of 60,000 or less.

  The present invention also provides the water-soluble β-glucan, which is an enzyme that catalyzes a reaction in which the enzyme to be added and acted on hydrolyzes between β-D-glucose bonds.

An effect of the present invention is to provide a β-glucan and a β-glucan complex having an excellent bioregulatory function and physiologically active function. In addition, it is safe to be used in the production of food, pharmaceuticals, and cosmetics, it is very easy to handle by suppressing problems such as thickening during production and use, and it is easy to powder and concentrate, making it the target formulation It is possible to provide a low molecular β-glucan whose amount is not substantially limited.
In addition, according to the present invention, it is excellent in solubility in water, stable even when stored in an aqueous solution without causing changes over time such as precipitation and turbidity, and does not precipitate even after repeated freezing / thawing. In addition, it is possible to provide a water-soluble β-glucan having excellent solubility.

Hereinafter, the present invention will be described in detail.
First, the β-glucan of the present invention will be described.

  The β-glucan of the present invention is a β-glucan derived from a microorganism, a β-glucan derived from a cereal, or a β-glucan derived from a basidiomycete.

  Next, the microorganism-derived β-glucan used in the present invention will be described.

  Since microorganisms themselves contain a large amount of β-glucan on their cell walls, microorganism-derived β-glucan is obtained by inoculating microorganisms into their respective growth media and growing the cells. The cultured cell wall residue obtained by disrupting the cultured cells and removing the contents as they are can be used. In addition, any of the cultured cells or the β-glucan extracted from the cultured cell wall residue can be used as it is, or purified of the extracted β-glucan. It is also possible to utilize β-glucan secreted and produced outside the cells by culturing microorganisms. In this case, the culture solution after completion of the culture can be isolated or purified from the culture solution as it is. Β-glucan can be used.

  Among these, when cultured cells obtained by inoculating microorganisms in each growth medium and growing the cells are used as they are, the cell contents are mixed, so the cultured cells are crushed and the contents are removed. It is preferable to use the cultured cell wall residue obtained in the above, 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 further to the outside of the cells. It is most preferable to use a secreted and produced β-glucan together with the culture solution or purified from the culture solution.

  As microorganisms suitable for obtaining the β-glucan, microorganisms that have been conventionally used for food are highly safe and suitable. That is, yeast, lactic acid bacteria, natto bacteria, acetic acid bacteria, koji molds, algae such as chlorella and spirulina, microorganisms belonging to the genus Aureobasidium, and the like. These can use the said microorganisms isolate | separated from the environment (for example, foodstuff, 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 UV treatment, 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, sparkling sake, shochu, sake, wine, whiskey, etc., for example, Saccharomyces cerevisiae (S. cerevisiae), Saccharomyces salmon (S.sake), Saccharomyces rosei (S.rosei), Saccharomyces ruxii (S.rouxii), Saccharomyces bisporus, Saccharomyces bailii (S.baillii), Saccharomyces bay Eggplant (S.bayanus), Saccharomyces capenisis, etc., and the genus Syzosaccharomyces, such as S. pombe, Torulopsis, such as Torropsis etkersi (T.etchelsii), Truropsis versatilis (T.versatilis), Truro T. holmii, Hanseniaspora, Hansenula, eg, H. subpelliculosa, Debaryomyces, eg, Debaryomyces Hanseni (D. hansenii), Saccharomycopsis genus For example, Saccharomycopsis fibrigera, Saccharomycodes genus, Pichia genus, Pachysolen genus, microorganism Examples include yeast of the genus Candida used for protein production, such as C. utilis, C. milleri, C. tropicalis, C. maltosa and C. lipolytica. In addition, Rhodotorula yeast.

  Examples of the lactic acid bacteria include genus Lactobacillus, Bifidobacterium, cocci, Leuconostoc, Pediococcus, Streptococcus, and Lactococcus. ) Lactic acid bacteria are usually used, but other genus Enterococcus genus, Vagococcus genus, Carnobacterium genus, Aerococcus genus, Tetragenococcus genus lactic acid bacteria 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 Mecenteroides (Ln.mesenteroides), Leuconostococcus (Ln.ocnos), Pediococcus acidilactici (Pediococcus acidilactici), Pediococcus cerevisiae (P.cerevisiae), Pediococcus spentaseseus (P.pentosaceus) One kind or two or more kinds of conventionally used lactic acid bacteria can be used. These may be used alone or in combination of two or more. Further, the culture of lactic acid bacteria belonging to the genus Bifidobacterium and the culture of other lactic acid bacteria may be performed separately, and these may be mixed.

  The microorganism belonging to the genus Aureobasidium may be any microorganism as long as it is a strain that produces a glucose polymer having a β bond outside the bacterial cell by culturing the microorganism, and examples thereof include Aureobasidium. A strain of Aureobasidium pullulans, specifically, IFO4464, IFO4466, IFO6353, IFO7757, ATCC9348, ATCC3092, ATCC42023, ATCC433023, FERM BP-8391 and the like can be used. 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 UV treatment, chemical treatment with nitrosoguanidine, ethidium bromide, ethyl methanesulfonate, sodium nitrite, and the like.

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

  As a method for isolating the cultured cell wall residue of the above microorganisms as β-glucan, a suitable amount of solvent is added to the cultured microorganisms, bacterial cells, and bacterial strains, and a part of the cell wall is added by autolysis or addition of hydrolase. In this method, the cultured cell wall residue is isolated as β-glucan by destroying the contents and allowing the contents to flow away and recovering the remaining components. Also, a method of obtaining β-glucan by damaging microorganisms or basidiomycetous cells by physical force such as French press or ultrasonic crusher, destroying part of the cells, removing the contents, and collecting the residue There is also.

  The extraction method of β-glucan is not particularly limited, and it may be extracted by adding an extraction solvent to microorganisms that are extraction raw materials. 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 manufacturing method, any form, and any purity can be used. Of course, it is also possible to use a mixture of extracted components other than β-glucan. In the present invention, all of these are referred to as β-glucan extracted from microorganisms.

  Furthermore, the method for extracting β-glucan from microorganisms will be described. Β-glucan used in the present invention can be dissolved in a solvent such as water as a water-soluble polymer, for example, by adjusting cell wall components of the bacterial cells, Using freeze-dried powder, water, warm water, hot water or salt solution, and acid, alkaline aqueous solution, organic solvent, etc., in a solvent 2 to 100 times the amount of powder for any time, at any temperature Can be extracted. Furthermore, β-glucan can be obtained by solid-liquid separation of the extract. Among these, β-glucan extracted with water, hot water or hot water is preferable, and β-glucan extracted with water having a temperature of 90 ° C. or lower and 4 ° C. or higher is more preferable. Further, an extraction accelerator such as an enzyme solution may be added at the time of extraction to perform extraction while reducing the molecular weight.

  The β-glucan derived from the microorganism of the present invention has 1-2β-D-glucopyranose bond, 1-3β-D-glucopyranose bond, 1-4-4-β-D-glucopyranose bond, 1-6 β-glucan having at least two kinds of β-D-glucopyranose bonds is preferable, and β-glucan composed of 1-3-β-D-glucopyranose bond and 1-6-β-D-glucopyranose bond is particularly preferable, so-called β1-3 and 1-6 glucans are preferred.

  In addition, when the extract from microorganisms is used as it is without purification, or the extract obtained by pulverizing and solidifying only is used as it is, the purity of β-glucan in the component is 1 to It may be 100%, preferably 10 to 100%, more preferably 20 to 100%, and the higher the purity, the better.

  The β-glucan of the microorganisms of the present invention is a high-molecular body, and β-glucan having any weight average molecular weight can be used. However, since the water solubility increases as the molecular weight decreases, the molecular weight is 3 million or less, preferably 500,000. Hereinafter, 100,000 or less is more preferable. For example, β-glucan obtained by extraction from microorganisms may be reduced in molecular weight by a known method so as to improve water solubility, or β-glucan having a low molecular weight may be directly extracted.

  Next, β-glucan derived from grains will be described. As the cereals, gramineous plants are preferable. Examples of gramineous plants include cereals such as rice, wheat, corn, sorghum, millet, millet, millet, barley, oats (crown) and rye In particular, those derived from barley are preferred. In particular, β-glucan obtained by extraction from these grasses is preferred. In the present invention, β-glucan obtained by this extraction is also referred to as extracted β-glucan.

  The extraction method of β-glucan extracted from this gramineous plant is not particularly limited, and it may be extracted by adding an extraction solvent to the gramineous plant as an extraction raw material. Also, the purity of the extract itself after solid-liquid separation, or a liquid or solid obtained by concentrating β-glucan extracted from the extract by a known method, or purified from the extract by a known method Any of the liquids, solids, etc. obtained by any manufacturing method, any form, and any purity can be used. Of course, a mixture of extracted components other than β-glucan may be used. In the present invention, these are all referred to as β-glucan extracted from the grass family.

  For extraction, the whole plant can be used as a raw material, but it is preferable to use seeds having a relatively high β-glucan content. Any of crushed powder (whole grain), cocoon, bran, malt, germ, and endosperm obtained in the grain refining process may be used. Preferably, it is an endosperm portion obtained by scouring whole grains and grains of barley and oats from the outer periphery, and rice bran, rice bran, wheat and corn bran and germs, and more preferably barley and oats. It is the endosperm part which refined the whole grain flour and grain of the kind from the outer peripheral part, and the cocoon which generate | occur | produces in that case.

  In addition, β-glucan extracted from gramineous plants has a 1-2β-D-glucopyranose bond, a 1-3β-D-glucopyranose bond, a 1-4β-D-glucopyranose bond, Β-glucan having at least two kinds of -6-β-D-glucopyranose bonds is preferable, and β-glucan consisting of 1-3-β-D-glucopyranose bonds and 1-4-β-D-glucopyranose bonds is preferable. So-called β1-3, 1-4 glucan is preferable.

  Explaining the method for extracting β-glucan from the grass family, β-glucan in the grass family can be dissolved as an aqueous solution as a water-soluble polymer. For example, water, warm water, hot water is added to the grain powder of the grass family. Alternatively, extraction can be performed at an arbitrary temperature for an arbitrary time using a salt solution, an acid, an alkaline aqueous solution, an organic solvent, etc., with a solvent 2 to 100 times the amount of the powder. Furthermore, 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 warm water having a temperature of 80 ° C. or lower and 4 ° C. or higher is more preferable. Further, an extraction accelerator or the like may be added during extraction.

  Specifically, as a method for obtaining high molecular weight β-glucan from barley, for example, a method of producing a waxy barley as a raw material by water extraction (Japanese Patent Publication No. 4-11197), or barley, oats A method for obtaining β-glucan having a weight average molecular weight of 100,000 to 1,000,000 by alkali extraction, neutralization and alcohol precipitation (Japanese Patent Publication No. 6-83652), and barley koji with a yield of 82% or less as a raw material And a method of extracting β-glucan with hot water at 80 to 90 ° C. (Japanese Patent Laid-Open No. 11-225706) and the like, and further reducing the molecular weight of β-glucan obtained by these production methods by a known method It can also be β-glucan. For example, as a method for reducing the molecular weight, any known hydrolysis reaction of polysaccharides can be used. For example, it is known that water-soluble polysaccharides are hydrolyzed by heating under pressure in the presence of an acid, and this can be used to reduce the molecular weight. Further, it is also effective to reduce the molecular weight using an enzymatic hydrolysis reaction. As the enzyme, 1,3-β glucanase or the like can be used. Furthermore, β-glucan obtained by direct extraction from raw material grains by methods such as WO98 / 13056 and JP-A-2002-97203 can also be used. Moreover, you may use the extraction promoter etc. which are described in Unexamined-Japanese-Patent No. 2002-105103.

  The grain-derived β-glucan used in the present invention is a high-molecular body, and β-glucan having any weight average molecular weight can be used. However, since the water solubility increases as the molecular weight decreases, the molecular weight is preferably 3 million or less, preferably It is preferably 500,000 or less, more preferably 100,000 or less. For example, β-glucan obtained by extraction from a gramineous plant may be reduced in molecular weight by a known method so as to improve water solubility, or β-glucan having a low molecular weight may be directly extracted.

  Next, β-glucan derived from basidiomycetes will be described.

  Basidiomycetes contain a large amount of β-glucan in the nuclei of fruit bodies and mycelia assembled in a lump, so that the fruit bodies and mycelia are finely pulverized, or the extract extracted from the pulverized product, or extracted Any of these can be used as a basidiomycete-derived β-glucan such as a product obtained by purifying β-glucan from a product. In addition, germination of basidiomycetous spores, inoculating mycelium in each growth medium and growing the bacterial cells, the cultured cells were obtained as they were, and the cultured cells were crushed and the contents were removed. Cultured cell wall residue can be used. In addition, any of the β-glucan extracted from the cultured cells or the cultured cell wall residue can be used as it is or purified from the extracted β-glucan. 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 is used as it is or separated and purified from the culture solution. β-glucan can be used as β-glucan derived from basidiomycetes.

  Among these, if the cultured cells obtained by inoculating spores and mycelium into each growth medium and growing the cells are used as they are, with the pulverized fruit bodies and mycelia being crushed, the cell contents However, it is preferable to use a cultured cell wall residue obtained by crushing the cultured cells and removing the contents, and further using the cultured cells or β-glucan extracted from the cultured cell wall residues as they are. Alternatively, it is more preferable to use after purification, and it is most preferable to use β-glucan secreted and produced outside the cells together with the culture solution or purified from the culture solution.

  As basidiomycetes, cultivars are most preferable, but basidiomycetes derived from basidiomycetes that are not used for commercial production may also be used. Examples include Agaricus blazei, Agaricus mushrooms, Agaricus mushrooms, Ezoharitake, Enokitake, Kanzotake, Mushroom, Kinugasatake, Agaricus, Salmonella mushroom, Sakerihi Toyotake, Sangohatake, Shitaketake, Shirotaketa Winter summer grass, sea cucumber, aratake, naratakemodoki, niboshimeji, nikawaurokotake, nikawaharitake, numerisugitake, numerisugitaketake, mushroom, oyster mushrooms, bullflower, owl, bamboo shoots, mushrooms, mushrooms, mushrooms , Mukitake, Murasakishiji, Yamadoritake, Yamabushitake, Willow Matsutake, Hanabiratake, Meshimako Etc. The.

  As a method for isolating the cultured cell wall residue of the above basidiomycetes as β-glucan, an appropriate amount of solvent is added to the cultured mycelium or the cultivated mycorrhiza or fruiting body, and autolysis or hydrolysis enzyme is added to the cell wall. A method of isolating the cultured cell wall residue as β-glucan by destroying a part and allowing the contents to flow away and recovering the residual components can be mentioned. There is also a method of obtaining β-glucan by damaging basidiomycete cells by physical force such as a French press or ultrasonic crusher, destroying a part thereof, removing the contents, and collecting the residue.

  The extraction method of β-glucan is not particularly limited, and it may be extracted by adding an extraction solvent to basidiomycetes serving as an extraction raw material. 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 manufacturing method, any form, and any purity can be used. Of course, it is also possible to use a mixture of extracted components other than β-glucan. In the present invention, all of these are referred to as β-glucan extracted from basidiomycetes.

  Furthermore, a method for extracting β-glucan from basidiomycetes will be described. Β-glucan used in the present invention can be dissolved in a solvent such as water as a water-soluble polymer, and is generally commercially available as basidiomycetes, for example. Mushrooms dried and pulverized powder, water, warm water, hot water or salt solution, further using acid, alkaline aqueous solution, organic solvent, etc., for 2 hours to 100 times the amount of solvent for the powder, It can be extracted at any temperature. Furthermore, β-glucan can be obtained by solid-liquid separation of the extract. Among these, β-glucan extracted with water, hot water or hot water is preferable, and β-glucan extracted with water having a temperature of 90 ° C. or lower and 4 ° C. or higher is more preferable. Furthermore, you may add extraction promoters, such as an enzyme solution, at the time of extraction.

  Β-glucan used in the present invention has 1-2β-D-glucopyranose bond, 1-3β-D-glucopyranose bond, 1-4-β-D-glucopyranose bond, 1-6-β. Β-glucan having at least two kinds of -D-glucopyranose bonds is preferable, and β-glucan consisting of 1-3-β-D-glucopyranose bond and 1-6-β-D-glucopyranose bond is particularly preferable, so-called β1 -3, 1-6 glucan is preferred.

  In addition, when the extract from basidiomycetes is used as it is without purification, or the extract is pulverized and solidified only as it is, the purity of β-glucan in the component is 1 to It may be 100%, preferably 10 to 100%, more preferably 20 to 100%, and the higher the purity, the better.

  The β-glucan derived from basidiomycetes used in the present invention is a high molecular body, and β-glucan having any weight average molecular weight can be used. However, since the water solubility increases with a decrease in molecular weight, the molecular weight is preferably 3 million or less, preferably Is less than 500,000, more preferably less than 100,000. For example, β-glucan obtained by extraction from basidiomycetes may be reduced in molecular weight by a known method so as to improve water solubility, or β-glucan having a low molecular weight may be directly extracted.

  The above-described β-glucan derived from the microorganisms of the present invention, β-glucan derived from cereals and β-glucan derived from basidiomycetes are bioregulatory functions and physiologically active functions such as lipid metabolism improving action, intestinal regulating action, and suppression of blood sugar level increase. It has an action, a cholesterol lowering action, a blood sugar level lowering action, an antitumor action, an immune enhancing action, an immunostimulating action, an intestinal immunity enhancing action, a skin immunity enhancing action, a lifestyle-related disease prevention / amelioration action and the like.

  Next, the β-glucan complex of the present invention will be described.

  The β-glucan derived from the microorganisms of the present invention, the β-glucan derived from cereals, and the β-glucan derived from basidiomycetes may be used singly, but by forming a β-glucan complex containing any two or more of them It produces a synergistic effect and becomes a material that is further excellent in biological regulation functionality, physiological activity function, and the like.

  In order to obtain a β-glucan complex, two or more types of β-glucan may be blended. For example, a complex of β-glucan derived from microorganisms and β-glucan derived from grains, β-glucan derived from microorganisms and β-derived from basidiomycetes Complex of glucan, complex of β-glucan derived from grain and β-glucan derived from basidiomycetes, complex of β-glucan derived from microorganisms, β-glucan derived from grains and β-glucan derived from basidiomycetes, and a complex of β1-3, 1-6 glucan and β1-3, 1-4 glucan.

  A method of blending two or more kinds of β-glucan to obtain a β-glucan complex is not particularly limited, and for example, β-glucan in a state of solid, powder, liquid, slurry, aqueous solution, solution or the like may be mixed. Stirring may be performed during mixing, and mixing may be performed at a constant temperature. (Stirring) The mixing temperature is preferably 30 to 130 ° C, more preferably 50 to 70 ° C, or 110 to 130 ° C. Moreover, it is also preferable to leave still for a fixed time (for example, 10 minutes-1 hour) after completion | finish of stirring and mixing. The order of adding each β-glucan is not particularly limited, and a processed product of β-glucan may be mixed.

  The blending ratio of β-glucan is not particularly limited. For example, the ratio of β1-3,1-6 glucan to β1-3,1-4 glucan is 0.1: 99.9 to 99.9: 0.1. 10-90: 90-10 are more preferable.

  The β-glucan complex of the present invention may be mixed with α-glucan as an optional component.

  The β-glucan complex of the present invention has a bioregulatory function and a physiologically active function, for example, lipid metabolism improving action, intestinal regulating action, blood sugar level rise inhibiting action, cholesterol lowering action, blood sugar level lowering action, antitumor action, immune enhancement It has an action, an immunostimulatory action, an intestinal immunity enhancing action, a skin immunity enhancing action, a lifestyle-related disease prevention / amelioration action, etc., and further has an excellent emulsifying action, taste and taste.

  Next, a method for producing a low molecular weight β-glucan will be described. The low molecular weight β-glucan of the present invention is water-soluble and is also referred to as a water-soluble β-glucan.

  First, a method for producing a low molecular weight β-glucan derived from microorganisms having a step of allowing a hydrolase to act is described.

  The microorganisms used in the method for producing a low molecular weight β-glucan derived from microorganisms of the present invention may be any of the microorganisms capable of obtaining the β-glucan, and in particular, microorganisms belonging to the genus Aureobasidium Is preferred. Hereinafter, a case where a microorganism belonging to the genus Aureobasidium is used will be described as an example.

  The method for producing a low molecular weight β-glucan derived from microorganisms of the present invention may include a step of allowing a hydrolase to act. That is, hydrolyzing enzyme may be allowed to act while producing β-glucan by a microorganism, that is, during the production process of β-glucan, or the resulting microorganism-derived β-glucan is allowed to act on hydrolyzing enzyme to reduce the molecular weight. Also good.

  Explaining the production process of β-glucan, a microorganism strain belonging to the genus Aureobasidium may be allowed to act on a medium in which the microorganism can grow to produce β-glucan in a medium outside the cells. In addition, a cultured microbial cell obtained by culturing a microbial strain belonging to the genus Aureobasidium is isolated, and the cultured microbial cell is allowed to act on a solution or medium containing a saccharide which is a substrate for β-glucan, so that Β-glucan may be produced. However, since there may be cases where secreted β-glucan prepared for secretion outside the cell body is encapsulated in the cell body, β-glucan prepared and accumulated in the cell body for secretion in this way is also the present invention. Is a β-glucan that is secreted outside the cell.

  In the method for producing a low molecular weight β-glucan according to the present invention, the medium for culturing the microorganism strain according to the present invention contains nutrient sources (carbon source, nitrogen source, inorganic salts) that can be normally used by microorganisms belonging to the genus Aureobasidium. Further, if necessary, a normal medium containing an organic nutrient can be used. As these media, various synthetic media, semi-synthetic media, natural media, etc. can be used, and the microorganisms are good. It may be performed under aerobic conditions such as aeration stirring and shaking under conditions that allow it to grow.

  In addition, when the cultured microbial cells obtained by culturing the microbial strain in the present invention are isolated and a low molecular weight β-glucan is produced using the cultured microbial cells or an extract of the cultured microbial cells as a catalyst, Add a saccharide solution or medium as a substrate to the suspension of the bacterial cells, cultured bacterial cell preparations or treated cultured bacterial cells, determine appropriate conditions in a known fermentation production method, Glucan can be produced. Examples of the cultured cell preparation include cell disrupted liquid obtained by homogenizing the cultured cell, and the cultured cell processed product is obtained by subjecting the cultured cell or cell disrupted solution to ion exchange in an alginate gel. Examples include immobilized cells immobilized on resin, ceramic, chitosan, and the like.

  The β-glucan secreted and produced outside the cells obtained by the above method can be modified (lower molecular weight) by enzymatic treatment before, during or after separation of the cells. That is, β-glucan to be subjected to enzyme treatment is obtained by culturing a culture solution containing high molecular weight β-glucan obtained by culturing the microbial strain of the present invention, a dried product of the culture solution, and β-glucan isolated from the culture solution. There are no particular problems whether it contains a solution or a dried product, and it contains β-glucan alone or a component other than β-glucan, and the state of the solution or dried product is not particularly limited. What is necessary is just to melt | dissolve a dried material suitably and to make an enzyme act. However, as described above, extraction of β-glucan immediately after production of microorganisms belonging to the genus Aureobasidium is very difficult industrially due to viscosity and thixotropy, and it is performed after or with enzyme action. It is preferable. Further, the step of secreting and producing β-glucan outside the microbial cells and the enzyme treatment in the present invention can be performed in parallel.

  The method for separating the cells may be according to a conventional method. Specifically, a method for separating and removing solids such as bacterial cells from the culture solution by centrifugation, filtration, etc., a method for removing impurities and salts with activated carbon, ion exchange resin, etc., or water, a salt solution, an acid aqueous solution, Various known purification means such as an extraction method using one or two or more mixed solvents such as an alkaline aqueous solution and an organic solvent can be selected and combined. Furthermore, for example, adsorption / elution to hydrophobic resin, solvent precipitation using ethanol, methanol, ethyl acetate, n-butanol, etc., column method using silica gel, etc., or thin layer chromatography, for fractionation using reverse phase column Separation and purification can be performed by using high performance liquid chromatography alone or in combination as appropriate, and optionally using it repeatedly. In addition, when β-glucan accumulated in cells or β-glucan constituting the cell wall is to be extracted, a part of the cell wall of the microbial cell body is destroyed in advance by autolysis or addition of hydrolase. Discharge the contents, destroy a part of microbial cells by physical force such as French press or ultrasonic crusher, let the contents flow out, add cellulase, glucanase, etc. to partially decompose the cell wall It is also possible to keep it.

  In addition, before and after separating β-glucan secreted and produced outside the cells by the above method, the cells may be sterilized. The sterilization temperature is not particularly limited as long as it is a temperature at which the cells die, but is preferably 50 ° C. or higher, more preferably 60 ° C., and most preferably 80 ° C. or higher. Moreover, the secretory β-glucan prepared in the cells can be extracted with hot water at a higher temperature, for example, at 90 ° C. or higher or 121 ° C. under pressure. The sterilization time and hot water extraction time can be set arbitrarily, but preferably 10 minutes or more and 8 hours or less, more preferably 15 minutes or more and 6 hours or less, and most preferably 30 minutes or more and 2 hours or less. Mixing is suppressed and β-glucan is not deteriorated, which is preferable.

  The enzyme treatment in the present invention is not particularly limited as long as the enzyme can be brought into contact with the β-glucan obtained by the step of producing β-glucan by the method. For example, the enzyme is added to β-glucan and stirred. What is necessary is just to stir with a blade | wing or a shaker. Of course, the enzyme may be brought into contact with β-glucan already obtained. In addition, it is more effective and preferable to perform ultrasonic treatment, homogenization treatment, ultra-high pressure treatment, heat treatment, etc. in any one or two or more stages after the enzyme treatment, before the enzyme treatment, during the enzyme treatment. .

  The method for isolating β-glucan having a low molecular weight by the enzyme treatment in the present invention is not particularly limited, and one or two or more mixed solvents such as water, salt solution, acid aqueous solution, alkaline aqueous solution, organic solvent, etc. Can be used. In particular, precipitation by an organic solvent and isolation by a molecular sieve (gel filtration) column are effective.

  As the enzyme used for the enzyme treatment of the present invention, an enzyme having an activity of hydrolyzing a β-D-glucoside bond can be used. However, an exo-type enzyme that sequentially hydrolyzes from the end of a polysaccharide chain produces a monosaccharide. Therefore, it is not suitable for the purpose of the present invention, and an enzyme that hydrolyzes an end type that hydrolyzes bonds other than the ends of the polysaccharide chain is preferable, and Endo-1,3-β-D-glucanase (EC 3.2. 1.6) is particularly preferred. Moreover, since the β-glucan produced by the microbial strain in the present invention has many branched structures, in addition to the enzyme that degrades the 1,3-β-D-glucoside bond, the 1,2-β-D-glucoside bond is degraded. Enzyme (EC 3.2.1.71), enzyme that breaks down 1,4-β-D-glucoside bond (EC 3.2.1.4), enzyme that breaks down 1,6-β-D-glucoside bond It is more effective when used together with (EC 3.2.1.75). In addition, an extract of a bacterial cell obtained by culturing a microorganism that produces these enzymes or a component obtained in a medium can be used as it is or purified as an enzyme in the present invention. As the microorganism, Aspergillus can be used. Aspergillus genus, Humicola genus, Rhizopus genus, Trichoderma genus, Acremonium genus fungus, Pycnoporus genus, Corticium genus, Irpex Genus, basidiomycetes belonging to the genus Rhizoctonia, Arthrobacter genus, Bacillus genus, Pseudomonas genus bacteria, Actinomyces genus, Streptomyces genus , Actinomycetes belonging to the genus Penicillium, Saccharomyces ( Yeast belonging to the genus Saccharomyces and Trichosporon is preferred, and in particular, Aspergillus aculeatus, Aspergillus niger, Aspergillus awamori, Aspergillus aceli Asperus alli・ Olysee (Aspergillus oryzae), Aspergillus usamii, Aspergillus japonicus, Aspergillus pulverulentus, Humicola insolens (Humicola insolens) (Rhizopus oryzae), Trichoderma harzianum, Trichoderma longibrachiatum, Trichodma longibrachiatum erma viride, Trichoderma insolens, Trichoderma koningii, Trichoderma reesei, Acremonium cellulolyticus, Pycnoporus coccus (Irpex lacteus), Rhizoctonia solani, Bacillus subtilis, Bacillus circulans, Bacillus amyloliquefaciens, Pudomonis pudois -Penicillium emersonii and Penicillium funicolosum are preferred.

  In addition, it is more effective when used in combination with one or more kinds of enzymes or substances containing enzymes that are commercially available or produced by the above microorganisms.

  The conditions for the enzyme treatment in the present invention may be appropriately determined depending on the properties of the enzyme used. For example, 0.001 to 10% of the enzyme is added to β-glucan and 5 to 5 at pH 2 to 9 is used. It is 10 minutes to 48 hours at 60 ° C., and particularly preferred conditions are 0.01 to 1% enzyme added to β-glucan, and pH 1 to 24 at 25 to 45 ° C. for 1 to 24 hours.

  Further, the treatment with an acid can be performed before and / or after the enzyme treatment. Examples of acids include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, and polyphosphoric acid, organic acids such as citric acid, lactic acid, succinic acid, acetic acid, malic acid, and gluconic acid, and salts thereof, and the target pH is What can be achieved, further, acidic electrolyzed water and the like can be mentioned. Among them, organic acids are preferable, and citric acid, acetic acid and lactic acid are particularly preferable. Examples of the alkali include sodium, potassium, magnesium, calcium and the like, alkali metal and alkaline earth metal hydroxides and salts, those capable of achieving the target pH with ammonia, and alkaline electrolyzed water.

  An immobilized enzyme can be used for the enzyme treatment in the present invention. Types of carriers that can be used for immobilization include solid carriers such as cation or anion exchange resin, chitosan, cellulose, ceramic, hydroxyapatite, activated carbon, porous glass, alumina, silica gel, polyacrylamide, κ- Any type of water-insoluble carrier such as a gelling agent such as carrageenan or alginic acid may be used, but a water-insoluble porous carrier processed into a porous material is particularly preferable. Two or more of these carriers can be used in combination. The shape of the carrier is not particularly limited, but a bead shape is preferable. Also, any size can be used.

  As a method for preparing the immobilized enzyme, any appropriate method can be used depending on the nature of the carrier used. Specifically, a carrier binding method such as a covalent bond method, an ionic bond method, a physical adsorption method, or a crosslinking method can be used. Law and inclusion law. That is, in the case of immobilization by the carrier binding method, the method is not particularly limited as long as the activated carrier and the enzyme solution can be brought into contact with each other as necessary, and a method of mixing and stirring the simple substance and the enzyme solution, Alternatively, a method of passing an enzyme solution through a single unit packed in a column can be used. In the case of immobilization by the crosslinking method, a polyfunctional crosslinking agent such as glutaraldehyde may be added to the carrier before mixing the enzyme solution, simultaneously with the enzyme solution, or after mixing the enzyme solution. In the case of immobilization by the entrapment method, a gelling agent such as polyacrylamide, κ-carrageenan or alginic acid and an enzyme solution are mixed, gelled by a predetermined method, and processed into a bead-like shape as necessary. For the purpose of using β-glucan modified by immobilized enzyme treatment for the human body such as foods, pharmaceuticals, cosmetics, etc. Those fixed by the method are preferred.

  The low molecular weight β-glucan derived from the microorganism of the present invention has a bioregulatory function and a physiological activity function, for example, a lipid metabolism improving action, an intestinal regulating action, a blood sugar level rise inhibiting action, a cholesterol lowering action, a blood sugar level lowering action, an antitumor action It has an immune enhancing action, an immunostimulating action, an intestinal immunity enhancing action, a skin immunity enhancing action, a lifestyle-related disease prevention / amelioration action, etc., and further has an excellent emulsifying action, taste and taste.

  Next, a method for producing a low molecular weight β-glucan derived from basidiomycetes having a step of allowing a hydrolase to act will be described.

  The method for producing a low molecular weight β-glucan derived from a basidiomycete of the present invention may include a step of allowing a hydrolase to act. That is, a hydrolase may be allowed to act while producing β-glucan by basidiomycetes, that is, during the production process of β-glucan, or a hydrolase is allowed to act on the obtained β-glucan derived from basidiomycetes. May be used. Further, it may be allowed to act while extracting β-glucan.

  Basidiomycetes contain a large amount of β-glucan in the nuclei of fruit bodies and mycelia assembled in a lump, so that the fruit bodies and mycelia are finely pulverized, or the extract extracted from the pulverized product, or extracted Any product such as a product obtained by purifying β-glucan from a product can be used. In addition, germination of basidiomycetous spores, inoculating mycelium in each growth medium and growing the bacterial cells, the cultured cells were obtained as they were, and the cultured cells were crushed and the contents were removed. Cultured cell wall residue can be used. In addition, any of the cultured cells or the β-glucan extracted from the cultured cell wall residue can be used as it is, or purified of the extracted β-glucan. 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 is used as it is, during the culture, or from the culture solution. Separated and purified β-glucan can be used.

  Among these, if the cultured cells obtained by inoculating spores and mycelium into each growth medium and growing the cells are used as they are, with the pulverized fruit bodies and mycelia being crushed, the cell contents However, it is preferable to use a cultured cell wall residue obtained by crushing the cultured cells and removing the contents, and further using the cultured cells or β-glucan extracted from the cultured cell wall residues as they are. Alternatively, it can be used during extraction or after purification. Furthermore, β-glucan secreted and produced outside the cells can be used together with the culture solution, during secretion production, or purified from the culture solution.

  The conditions for enzyme treatment, the type of enzyme, etc. may be the same as in the method for producing a low molecular weight β-glucan derived from the microorganisms.

  Next, a method for producing a grain-derived low molecular weight β-glucan having a step of allowing a hydrolase to act is described.

  First, grain and / or grain-derived β-glucan that can be used as a raw material in the production method of the present invention will be described.

  As the cereal, cereals containing hemicellulose, particularly grasses are preferred, and specifically, barley, oats, barley bran, wheat germ, rice bran, corn hulls, etc. A relatively large amount of barley, oats, barley bran, oat pearl barley, barley hulls, and oat hulls are preferred.

  In the production method of the present invention, β-glucan derived from cereal can be used as a raw material. Β-glucan derived from cereal is β-glucan obtained by operations such as extraction and fractionation from cereal, and if β-glucan is contained, purified product, crude product, extract, concentrate, solution, It may be any of powder, liquid, gel, and the like, and may be a mixture in which other components contained in the cereal are mixed, all of which are referred to as cereal-derived β-glucan in the present invention.

  In the present invention, it is preferable to use β-glucan obtained by an operation such as extraction as a raw material, and it is preferable to use a raw material that has been treated in advance so that the content of β-glucan is increased. And more preferable. The β-glucan derived from cereal used as a raw material is particularly preferably an extract or fraction obtained by concentrating β-glucan than the grain, and the higher the β-glucan content, the better. In addition, as a raw material, the proportion of β-glucan is 7% by weight or more, preferably 30% by weight or more, more preferably 50% by weight or more in the grain and / or grain-derived β-glucan having a water content of 10% by weight or less. It is better to use raw materials. In the present invention, fractions, concentrates and extracts with an increased β-glucan content are also included in the β-glucan derived from grains. In addition, an extract or a fraction obtained by intentionally concentrating β-glucan and, as a result, enriching β-glucan, and other processed products can be used as a raw material. In particular, it is preferable to use an extract or fraction in which β-glucan is concentrated at a high concentration.

  Next, the enzyme used in the present invention will be described.

  As the enzyme used in the present invention, an enzyme having an activity of hydrolyzing β-glucan, that is, an enzyme having an activity of hydrolyzing a bond between β-D-glucose is preferably used. Specific examples include cellulase, β-glucanase, 1,3-β-D-glucanase, 1,4-β-D-glucanase, 1,6-β-D-glucanase, and complex enzymes thereof. In addition, commercially available enzyme preparations include enzymes and enzyme preparations that have these activities but have been developed for the purpose of degrading polysaccharides other than β-glucan, such as pectinase, xylanase, hemicellulase, cell wall lytic enzyme, and extract viscosity. Β, which is substantially derived from the above-mentioned grains, such as enzymes showing a reducing action, enzyme preparations for preventing sedimentation of fruit juice components, enzymes used for the purpose of reducing the viscosity of beer and fruit juice, and for the purpose of accelerating the filtration rate Any enzyme or enzyme preparation capable of reducing the molecular weight of glucan and preparing the water-soluble β-glucan of the present invention can be used. Commercially available enzymes that can be used in the present invention include, for example, lichenase (manufactured by Nippon Biocon), celex (manufactured by Novozyme), grainase (manufactured by Daiwa Kasei Kogyo), finizyme (manufactured by Novozyme), celeclast (novozyme). For example). Particularly preferred enzymes are lichenase, cerex mix and graynase.

  Moreover, at the time of reaction with the said enzyme, 1 or more types selected from alpha amylase, proteolytic enzyme, and glucoamylase can be added, and preparation of the water-soluble beta glucan of this invention can be accelerated | stimulated. α-amylase hydrolyzes α1-4 glycosidic bonds such as gelatinized starch and glycogen at any position to produce dextrin, oligosaccharide and glucose as degradation products. Glycoamylase is a substance that degrades α1-4 glycosidic bonds of soluble starch sequentially in glucose units from the non-reducing end. The proteolytic enzyme can be used to promote the preparation of the water-soluble β-glucan of the present invention by degrading soluble proteins extracted simultaneously with β-glucan and peptides complexed with β-glucan. These enzymes that carry out the accelerating reaction may be added together with the enzyme having the activity of hydrolyzing β-glucan, or may be used for concentrating, extracting, and fractionating β-glucan derived from cereal. May be used sometimes.

  Next, the reaction conditions for the enzyme will be described.

In the present invention, β-glucan is reduced in molecular weight by adding the above enzyme to cereal and / or cereal-derived β-glucan to act, thereby obtaining water-soluble β-glucan.
Any enzyme reaction conditions in the present invention can be used as long as they are used for ordinary enzyme reactions. In order to maintain the enzyme activity optimally, the temperature is preferably 80 ° C. or lower and 10 ° C. or higher. Depending on the characteristics of the enzyme, it may be preferable to perform the treatment at 80 ° C. or higher and 10 ° C. or lower. The pH is preferably near neutral, and is usually pH 10 to pH 4, preferably pH 9 to pH 5, and more preferably pH 8 to pH 6. In order to maintain the stability of the enzyme and β-glucan in the enzyme reaction, salts, acids and alkalis can be added and used as necessary.

  When the enzyme is allowed to act, 10 to 2000 parts by weight of water and 0.001 to 20 parts by weight of enzyme are added to 100 parts by weight of cereal and / or β-glucan derived from cereal, and 0.5 to 24 hours, preferably It is desirable to carry out the reaction with stirring for 1 to 12 hours, more preferably 1.5 to 6 hours. The amount of water used is more preferably 50 to 1000 parts by weight, and most preferably 100 to 500 parts by weight.

  β-glucan does not necessarily have to be completely dissolved in the enzyme reaction solution, and the solubility increases with the progress of the enzyme reaction and dissolves in the reaction solution. The state may be a completely dissolved product, a slurry state, a precipitated state, or the like, and the β-glucan solution does not need to be transparent and uniform. As long as the enzyme is not inactivated during the enzyme reaction, grains or grains-derived β-glucan (purified β-glucan solution, β-glucan solution in a slurry state, β-glucan powder, etc.) is added as appropriate. The enzyme reaction can be continued, or the enzyme reaction can be continued by continuously adding an enzyme or an enzyme solution.

  The enzyme reaction solution after the enzyme reaction for a certain time can be used as it is as a composition / mixture containing water-soluble β-glucan. In addition, it is possible to obtain a water-soluble β-glucan or a composition / mixture containing water-soluble β-glucan by adding steps such as solid-liquid separation and purification as necessary to the enzyme reaction solution after the enzyme reaction for a certain period of time. . The enzyme deactivation operation may be performed at any time point after the end of the enzyme reaction. A conventional method can be used as the deactivation method, and heat treatment is preferred. In addition, it is preferable to use an immobilized enzyme as the enzyme, and it is more preferable that the immobilized enzyme is allowed to act on cereals and / or cereal-derived β-glucan so that the enzyme is not mixed in the water-soluble β-glucan. .

  The low molecular weight (water-soluble) β-glucan obtained by the method for producing low-molecular weight (water-soluble) β-glucan of the present invention has good compoundability in foods, and is known to be a β-glucan derived from cereals that has been conventionally known. Since water solubility has been improved in comparison, it has a bioregulatory function such as lipid metabolism improving action, intestinal regulating action, blood sugar level increase suppression, etc., and has excellent emulsifying action, taste, taste, Taking advantage of these functions, emulsifiers, food modifiers, additives, humectants, oil and fat substitutes, and low calorie food materials are preferably used for foods, cosmetics, chemicals, and pharmaceuticals.

Next, the water-soluble β-glucan of the present invention obtained by adding an enzyme to cereal and / or cereal-derived β-glucan to reduce the molecular weight will be described.
The water-soluble β-glucan of the present invention is obtained by adding an enzyme to cereal and / or cereal-derived β-glucan to reduce the molecular weight, and the method for producing a low-molecular-weight β-glucan derived from cereal according to the present invention. Can be obtained. The water-soluble β-glucan referred to in the present invention is β-glucan that is soluble in water, and its solubility in water is higher than before by adding an enzyme and allowing it to act. In the present invention, the solubility in water means not only the solubility in water but also the dissolution without the need for excessive stirring, the dissolution at a low temperature, the one once dissolved, the temperature drops or refrigerated storage No gelation, precipitation, or turbidity, or once dissolved, gelation, precipitation or turbidity will not occur over time, gelation or precipitation even after repeated freezing and thawing The water-soluble β-glucan of the present invention is excellent in such solubility in water.

The water-soluble β-glucan of the present invention will be described in more detail from the viewpoint of solubility in water. The water-soluble β-glucan of the present invention preferably has a solubility in water at 25 ° C. of 1 mg / ml or more. More preferably, it is 10 mg / ml or more, and most preferably 50 mg / ml or more.
In addition, the water-soluble β-glucan of the present invention is dissolved in water at a concentration of 50 mg / ml or more at 25 ° C. to form an aqueous solution, and this aqueous solution is stored at room temperature, stored refrigerated (4 ° C.), or repeatedly frozen and thawed Furthermore, it is preferable that water solubility, such as gelatinization, precipitation, and turbidity, is improved.
The water-soluble β-glucan of the present invention is hydrated / dissolved when heated to 80 ° C. or higher, particularly 70 ° C. or higher, particularly 60 ° C. or higher and stirred and dissolved in water at a concentration of 30 mg / ml or higher. It is preferable that no precipitation occurs.
The water-soluble β-glucan of the present invention is heated to 80 ° C. or more, particularly 70 ° C. or more, particularly 60 ° C. or more, dissolved in water at a concentration of 50 mg / ml or more to prepare an aqueous solution, and this aqueous solution is stored at room temperature. It is preferable that water solubility such as gelation, precipitation, and turbidity is improved when refrigerated storage (4 ° C.) or when freezing and thawing are repeated.
In addition, when the water-soluble β-glucan of the present invention is added at a concentration of 50 mg / ml or more with respect to water, it dissolves even when the temperature of the water is 30 ° C. or lower, and no precipitation is observed. It is preferable that precipitation does not occur even when thawing is repeated.

  About the test method of solubility of water-soluble β-glucan in water, as described in Test Examples 1 to 4 described later, a method of visually confirming the gelation and dissolution state, and a method of measuring the absorbance of the solution at 660 nm In addition, general physical property tests and methods for investigating physicochemical properties, such as viscosity measurement, physical property measurement of gelled products (breaking strength measurement with rheometer, gel strength), gel melting test, etc. Any method may be used, such as a method for evaluating thickeners or gelling agents, or a method for testing the dissolved state of substances. As a comparative control in the test, β-glucan before the enzyme is added and allowed to act may be used, or it may be compared with a commercially available barley β-glucan standard product.

  The structure of the water-soluble β-glucan of the present invention is mainly composed of D-glucose, and the bond is mainly 1,3 bond and 1,4 bond, that is, glucose is β-1,3 bond or β It is a so-called 1,3,1,4-β-D-glucan that is bonded to a straight chain by -1,4 bonds, and the ratio (the former: the latter) is approximately 1: 2 to 1: 4. Of course, as long as it is obtained by adding an enzyme to cereal-derived and / or cereal-derived β-glucan, it may have β-1,2 bond, β-1,6 bond, etc. .

  The solubility of β-glucan correlates with the molecular weight, and the lower the molecular weight, the better. The water-soluble β-glucan of the present invention preferably has a weight average molecular weight of 60,000 or less, 400 or more, more preferably 40,000 or less, 400 or more, more preferably 30,000 or less, 400 or more, still more preferably 20,000 or less, 400 or more, most preferably Is 10,000 or less and 500 or more. It is preferable to lower the molecular weight of cereal-derived and / or cereal-derived β-glucan using an enzyme so that the weight average molecular weight is in these ranges.

  The water-soluble β-glucan of the present invention may be purified and used alone, or may be used together with other normal β-glucans. Alternatively, carbohydrates (starch, α-glucan), proteins, lipids, amino acids, peptides, nucleic acids, saccharides (monosaccharides, oligosaccharides), etc. that are usually contained in cereals and other components may be used.

  Next, the shape of the water-soluble β-glucan of the present invention will be described.

  The form of the water-soluble β-glucan of the present invention may be solid (powder), liquid, or slurry. After the enzyme is deactivated following the enzyme reaction, the reaction product may be used as it is, or the reaction product. Any shape can be used, such as a solidified product or a pulverized solidified product. In addition, a water-soluble β-glucan solution can be obtained by solid-liquid separation of the reaction product by an arbitrary method known as a solid-liquid separation method such as centrifugation, filtration separation, membrane separation, and natural sedimentation. Moreover, water can be added to the reaction product after completion of the enzyme reaction to extract water-soluble β-glucan with improved solubility.

  Furthermore, the water-soluble β-glucan solution can be concentrated by any method known as a liquid concentration method such as membrane concentration method, freeze concentration method, reduced pressure concentration method, salting out precipitation method, organic solvent precipitation method, In addition, it can be heated to evaporate water, solidify, or pulverize to form a composition or mixture containing water-soluble β-glucan or water-soluble β-glucan.

  One type of β-glucan is 1,3,1,6-β-D-glucan, and the 1,3,1,6-β-D-glucan can be obtained from microorganisms and basidiomycetes. . The 1,3,1,6-β-D-glucan, or β-glucan derived from microorganisms or basidiomycetes has physiological functions such as an immunopotentiating action and an antitumor activity action. When the water-soluble β-glucan of the present invention is used in combination with these β-glucans and used as a β-glucan complex, the physiological functions possessed by each are enhanced and a synergistic effect is produced. 6-β-D-glucan, or β-glucan derived from microorganisms or basidiomycetes is preferable because solubility in water is also improved. In order to obtain this β-glucan complex, 1,3,1,6-β-D-glucan or β-glucan derived from microorganisms or basidiomycetes and the water-soluble β-glucan of the present invention in an arbitrary ratio Can be mixed.

  The water-soluble β-glucan of the present invention has 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, antitumor action, Immune enhancement, immunostimulation, intestinal immunity enhancement, skin immunity enhancement, lifestyle-related disease prevention / amelioration, etc., excellent solubility in water, and emulsification, thickening, etc. It can be preferably used for foods, pharmaceuticals, cosmetics, feed, chemical products and the like. When used, the water-soluble β-glucan of the present invention may be used alone, or the water-soluble β-glucan of the present invention may be used in combination with other components. Since the water-soluble β-glucan of the present invention is particularly excellent in water solubility, it is easy to handle and easy to mix with other components.

  Β-glucan derived from microorganisms of the present invention, β-glucan derived from basidiomycetes, β-glucan derived from cereal, β-glucan complex, low-molecular-weight β-glucan obtained by the method for producing low-molecular-weight β-glucan derived from microorganisms, A low molecular weight β-glucan obtained by a method for producing a low molecular weight β-glucan derived from a basidiomycete, a low molecular weight β-glucan obtained by a method for producing a low-molecular weight β-glucan derived from a grain, and a water-soluble β-glucan, Excellent bioregulatory function and bioactive function, such as 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 Has immunity enhancing action, skin immunity enhancing action, lifestyle-related disease prevention / amelioration action, etc. Furthermore, it has emulsifiability, thickening, etc., and it is blended into foods, pharmaceuticals, cosmetics, feeds, chemical products, etc. Good Can be used properly.

  Examples of foods that can be used or blended include fat and oil foods, bakery products, confectionery, noodles, processed rice products, processed wheat products, processed corn products, processed products of beans (soybeans, red beans, etc.), cereals (buckwheat noodles, mushrooms, awa) , Millet) processed products, agricultural products (potato, sweet potato, taro, troroimo, etc.) processed products, dairy products, soups, beverages, seasonings, processed meat products, processed marine products, cooked / prepared foods, health foods, low-calorie 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.

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

  Examples of the bakery product include bread, white bread, sweet bread, sugar beet bread, French bread, croissant, pie, castella, sponge cake, butter cake, choux pastry, waffle, steamed bread, fermented sweet and the like.

  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, 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, monjayaki, 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 beverage, yuba, deep-fried oil, deep-fried chicken, cancer pounding, red bean paste, miso, various bean dishes, and the like.

  Examples of processed cereals (buckwheat, shrimp, sweet potato, millet) and processed agricultural products (potato, sweet potato, taro, troroimo, etc.) include buckwheat, potato snacks, potato chips, sweet potatoes, potato fries, various potato dishes, etc. 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 these may be added to these ingredients. Or you may add in the foodstuffs added directly to these soups 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 and the like.

  Examples of the above-mentioned seasonings include soy sauce, fish sauce (squid crab soy sauce, sardine soy sauce, salted 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, various meat dishes, and the like.

  Examples of the processed fishery product include kamaboko, deep-fried sweet potato, tsumire, and kneaded products.

  Examples of the health food or medicinal food include supplements, tablets, drinks, sports drinks and the like.

  In the case of blending into food, the blending amount may be determined according to the purpose. For example, it is preferably 80 to 0.01% by weight, particularly preferably 5 to 0.5% by weight based on the total amount of food.

  Examples of pharmaceuticals that can be used or formulated include lipid metabolism improving action, intestinal regulating action, blood sugar level rise inhibiting action, cholesterol lowering action, blood sugar level lowering action, antitumor action, immune enhancing action, immunostimulating action, intestinal immunity Drugs with potentiating action, skin immunity enhancing action, lifestyle-related disease prevention / amelioration action, etc., cancer prevention or treatment drugs, infection prevention or treatment drugs, skin treatment drugs, ointments, patches, nasal preparations , Ear drops, eye drops and the like. Furthermore, they are also useful as ointment bases, tablets, granules, excipients for powder formulations, internal use, and liquid dispersions.

  In the case of blending into a pharmaceutical product, the blending amount may be determined according to the purpose.

  Examples of feeds that can be used or formulated include livestock feed, pet feed, fish and shellfish feed, cultured fish feed, livestock therapeutic or preventive drugs, pet therapeutic or preventive drugs, fish and shellfish therapeutic drugs or Examples include preventive drugs, cultured fish treatment drugs or preventive drugs, and infectious disease preventive drugs.

  In the case of blending into feed, the blending amount may be determined according to the purpose. For example, it is preferably blended in an amount of 80 to 0.01% by weight, particularly 5 to 0.5% by weight based on the total amount of the feed.

  Β-glucan derived from microorganisms of the present invention, β-glucan derived from basidiomycetes, β-glucan derived from cereal, β-glucan complex, low-molecular-weight β-glucan obtained by the method for producing low-molecular-weight β-glucan derived from microorganisms, A low molecular weight β-glucan obtained by a method for producing a low molecular weight β-glucan derived from a basidiomycete, a low molecular weight β-glucan obtained by a method for producing a low molecular weight β-glucan derived from a grain, and a water-soluble β-glucan, When 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 and absorption control agents, anti-aging agents, antioxidants, blood circulation promoters, amino acids, peptides, lipids, polysaccharides, oligosaccharides, proteins, carbohydrates, dietary fibers, enzyme components, and the like.

  Specific examples of these physiologically active substances include polyunsaturated fatty acids (EPA, DHA) that optimize blood lipid levels, plant sterols that regulate serum cholesterol, and esterified products thereof, 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 And other 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, gaba, 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, Oregano, Catnip tea, Yarrow, Hypiscus, Echinesia and other herbs, vitamins, calcium-containing compounds such as calcium fortifiers, iron-containing compounds and other iron fortification , Mineral reinforcing agent containing essential minerals, more animals and plants extract, seaweed extract, bioactive ingredient natural ingredients such as crude drugs components and the like.

  As vitamins that can be blended, those listed as fortifiers in food additives can be used, such as vitamin A or beta carotene, vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, pantothenic acid , Niacin, biotin, vitamin C, vitamin D, vitamin E, vitamin K, derivatives thereof, oil-coated vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, pantothenic acid, niacin, biotin, vitamin C, coenzyme Q10 (Vitamin Q). Specific examples of vitamin derivatives include thiamine hydrochloride, thiamine sulfate, dibenzoylthiamine, dibenzoylthiamine hydrochloride, thiamine nitrate, thiamine lauryl sulfate, bisbenchamine as vitamin B1, and riboflavin butyrate as vitamin B2. And riboflavin 5′-phosphate sodium, pyridoxine hydrochloride as vitamin B6, L-ascorbic acid stearate, sodium L-ascorbate and the like 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.

  The edible molded product of the present invention may contain an essential mineral, and examples of the essential mineral include zinc, potassium, magnesium, manganese, phosphorus, sodium, selenium, iodine, and molybdenum. Examples of sources for supplying such essential minerals include wheat extract, wheat germ, young wheat leaf, chlorella, oyster meat extract, seawater concentrate, nuts, fish meal, liver powder, brown rice powder, and brewer's yeast.

  The biologically active components of naturally derived components such as animal and plant extract components, seaweed extract components and herbal medicine components that can be blended include 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, Oat Extract, Auren Extract, Barley Extract, Hypericum Extract, Oyster 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, kiwi extract, quinae , Cucumber extract, Guanosine, Gardenia extract, Kumazasa extract, Clara extract, Walnut extract, Grapefruit extract, Clematis extract, Chlorella extract, Mulberry extract, Gentian extract, Tea extract, Yeast extract, Burdock extract, 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, horseshoe extract, hawthorn extract, elderberry extract, horsetail Yarrow extract, mint extract, sage extract, mallow extract, sensu extract, soy extract, soybean extract, thyme extract, tea extract, clove extract, chigaya extract, chimpi extract, suzuki extract, eucalyptus extract, tonin extract, spruce extract, dokudami Extract, tomato extract, natto extract, carrot extract, garlic extract, wild rose extract, hibiscus extract, bacmond extract, parsley extract, honey, hamamelis extract, parietalia extract, toad extract, bisabolol, loquat extract, dandelion extract, burdock root extract Extract, butcher bloom extract, grape extract, placenta extract, propolis, loofah extract, safflower extract, peppermint Extract, body extract, button extract, hop extract, pine extract, maronier extract, mizubasho extract, mukuroji extract, melissa extract, peach extract, cornflower extract, eucalyptus extract, yukinoshita extract, yuzu extract, yokoinin extract, mugwort extract, lavender extract, 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, preservatives commonly used in foods , Fragrance, antibacterial agent, bactericidal agent, salt, solvent, chelating agent, neutralizing agent, acidulant, pH adjuster, preservative, buffering agent, fats and oils, processed fats, starch, wheat flour, buckwheat flour, flour, eggs, Components such as 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, coconut oil, palm kernel oil, hardened oil and other oils, various proteins, or wheat flour, rice flour, Azuki bean powder, soybean powder, wheat germ and other flours, soy protein, skim milk powder, casein, albumin, globulin, cheese and other dairy products and processed milk products, egg white, egg yolk and other egg processed products, sugar, glucose, maltose, Examples thereof include saccharides such as lactose, salts such as sodium chloride and calcium carbonate, and a mixture or composite material thereof may be used.

  Furthermore, β-glucan derived from the microorganisms of the present invention, β-glucan derived from basidiomycetes, β-glucan derived from cereals, β-glucan complex, and low-molecular-weight β-glucan obtained by the method for producing low-molecular-weight β-glucan derived from microorganisms , A low molecular weight β-glucan obtained by a method for producing a low molecular weight β-glucan derived from a basidiomycete, a low molecular weight β-glucan obtained by a method for producing a low molecular weight β-glucan derived from a grain, and a water-soluble β-glucan Examples of cosmetics that can be used or formulated include lotions, emulsions, skin milk, creams, ointments, external preparations, lotions, calamine lotions, sunscreen agents, suntan agents, after-shave lotions, pre-shave lotions, makeup bases, packs, Basic cosmetics such as cleansing products, facial cleansers, anti-acne cosmetics, and essences; foundations, white powder, eye shadows Make-up cosmetics such as C, eyeliner, eyebrow, teak, lipstick and nail 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, hay fever and allergy improving agents, antiallergic agents, rough skin prevention agents, skin disease improving agents, skin damage repairing agents, wrinkle protection agents, cold Examples include poultice, hot poultice, poultice, patch, scrub cosmetics, soap, tissue paper, body shampoo, hand soap, perfume, toothpaste, oral care products, mouthwash, gum massage cream, bathing agent, etc. .

  When blended in the cosmetic, 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.

  Substances that give physiological activity to the skin include, for example, naturally-occurring components such as animal and plant extract components, seaweed extract components and herbal components, 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, Oat extract, Ouren extract, barley extract, hypericum extract, Odrianthus extract, Dutch mustard extract, orange Extract, dried seawater, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, kawara mugwort extract, licorice extract, oil-soluble licorice extract, calcade extract, oyster Cucumber extract, Kiwi 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, 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, Citrus extract, Peonies extract, Ginger root extract, Birch extract, Horsetail extract, Kizuta extract, Hawthorn extract Elderberry extract, Achillea millefolium extract, Pepper mint extract, Sage extract, Zeni mushroom extract, Senkyu extract, Senbu extract, Soybean extract, Thymus 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, Fukitan Popo Extract , Fukinoto extract, Bukuryo extract, Butcher bloom extract, Grape extract, Placenta extract, Propolis, Loofah Extract, safflower extract, peppermint extract, body extract, button extract, hop extract, pine extract, maronier extract, honeysuckle extract, mugwort extract, melissa extract, peach extract, cornflower extract, eucalyptus extract, yukinoshita extract, yuzu extract, yokuinin 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, chondroitin sulfate sodium, collagen, elastin, chitin, chitosan Biopolymers such as hydrolyzed eggshell membranes; amino acid derivatives such as amino acids, sarcosine, N-methyl-L-serine; polyhydric alcohols or sugars such as ethyl glucose, sodium lactate, urea, sodium pyrrolidonecarboxylate, betaine, whey Other moisturizing ingredients; oily ingredients such as sphingolipids, ceramides, cholesterol, cholesterol derivatives, phospholipids; anti-inflammatory agents such as ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, guaiazulene, hydrocortisone; Mines A, B ₂ , B ₆ , D, vitamins such as calcium pantothenate, biotin, nicotinamide, vitamin E; active ingredients such as allantoin, diisopropylamine dichloroacetic acid, 4-aminomethylcyclohexanecarboxylic acid; carotenoids, flavonoids , Tannin, lignan, saponin and other antioxidants; α-hydroxy acid, β-hydroxy acid, mevalonic acid and other cell activators, γ-oryzanol and other blood circulation promoters, retinol, retinol derivatives and other wound healing agents; ascorbine Whitening agents such as acids, arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, glutathione; cephalanthin, licorice extract, red pepper tincture, hinokitiol, garlic iodide extract, pyridoxine hydrochloride, nicotinic acid, nicotine acid Conductor, 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 vanillyl amide, nonanoic acid vanillyl amide, piroctone olamine, glyceryl pentadecanoate, l-menthol, mononitroguaiacol, resorcin, γ-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormone, cantharis tincture, cyclosporine, zinc pyrithione, Hydrocortisone, minoxidil, polyoxyethylene sorbitan monostearate, mint oil, sasanishiki Examples include hair restorers such as extracts, 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. Examples include salts. 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, which are known substances and are described in JP-B-44-31237 and JP-B-54-21415. Examples of higher fatty acid esters of ascorbic acid include ascorbic acid-2-palmitic acid monoester, ascorbic acid-2,6-palmitic acid diester, ascorbic acid-2-stearic acid ester, and the like.

  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 above powders include dyes such as red 104, red 201, yellow 4, blue 1, black 401, rake dyes such as yellow 4 Al lake, yellow 203 Ba rake; nylon powder , Silk powder, urethane powder, Teflon (registered trademark) powder, silicone powder, polymethyl methacrylate powder, cellulose powder, silicone elastomer spherical powder, polyethylene powder, etc .; 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; Pearl pigment; barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate, silicate mug Metal salts such as Siumu; silica, inorganic powder such as alumina; bentonite, smectite, boron nitride, and the like. These powders have a spherical shape, a rod shape, a needle shape, a plate shape, an indefinite shape, a flake shape, a spindle shape, and the like.

  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.

  Oils include volatile and non-volatile oils and solvents and resins that are usually used in skin cosmetics, and may be liquids, pastes, or solids at room temperature, but liquids that are excellent in handling are preferred. Examples of oil agents 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, sorbitol, ethylene glycol, propylene Polyols and sugars such as glycol and polyethylene glycol; myristyl myristate, hexyl laurate, decyl oleate, isopropyl myristate, hexyl decyl dimethyloctanoate, glyceryl monostearate, diethyl phthalate, ethylene glycol monostearate, oxy Esters such as octyl stearate; Hydrocarbons such as liquid paraffin, petroleum jelly and squalane; Waxes such as lanolin, reduced lanolin and carnauba wax Include ethylene-alpha-olefin co-oligomer; mink oil, cacao butter, coconut oil, palm kernel oil, camellia oil, sesame oil, castor oil, oils such as olive.

  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, fluoroalcohol and the like can be mentioned.

  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. .

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

  Examples of 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 substituted acrylamide type Polymer, polyacrylamide, cationic polymer such as cationized gar gum, dimethylacrylammonium polymer, acrylic acid / methacrylic acid acrylic copolymer, POE / POP copolymer, polyvinyl alcohol, pullulan, agar, gelatin, tamarind seed polysaccharide, Xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, gar gum, gum arabic, crystalline cellulose, arabinogalactan, karaya gum, gum tragacanth, algin , 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 the ultraviolet protective agent (including organic and inorganic types, 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-methylbenzidine 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.

  Further, β-glucan derived from microorganisms of the present invention, β-glucan derived from basidiomycetes, β-glucan derived from cereals, β-glucan complex, low-molecular-weight β obtained by the method for producing low-molecular-weight β-glucan derived from microorganisms Glucan, low molecular weight β-glucan obtained by the method for producing low molecular weight β-glucan derived from basidiomycetes, low molecular weight β-glucan obtained by the method for producing low molecular weight β-glucan derived from cereal, and water-soluble of the present invention Examples of chemicals that can use or be incorporated with β-glucan include medical detergents, clothing detergents, contact lens preservatives, contact lens cleaning liquids, disinfectants, soaps, hygiene products, medical supplies, toiletries, thickeners, Examples thereof include a viscosity modifier.

  When blended into a chemical product, the blending amount may be determined according to the purpose. For example, it may be blended in an amount of 100 to 0.01% by weight, particularly 50 to 0.01% by weight based on the total amount of the chemical product. preferable.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but these do not limit the present invention. “Parts” and “%” are based on weight unless otherwise specified.

Example 1 Production Example 1 of Microbial Derived β-Glucan
Aureobasidium pullulans IFO-6353 strain was inoculated into YM medium and cultured at 26 ° C. for 3 days to obtain 300 mL seed culture solution. Into a 5L jar fermenter (Maruhishi Bioengineer) equipped with a full zone wing, put 3L of Kuzapek medium and 300g of sucrose, inoculate 100 mL of seed culture solution after sterilization and cooling, and incubate at 26 ° C for 72 hours. Culture was performed to obtain 3 L of a culture solution. The culture solution was sterilized by heating at 80 ° C. for 30 minutes to obtain a culture solution.

  To 1 L of the main culture solution, 10 g of cellulase preparation cellulase A “Amano” 3 (Amanoenzyme) was added and stirred at 40 ° C. for 20 hours, followed by enzyme treatment to obtain a low molecular weight β-glucan aqueous solution.

Example 2 Production Example 2 of Microbe-Derived Low Molecular Weight β-Glucan
Culture was performed in the same manner as in Example 1 except that the strain was Aureobasidium pullulans ADK-34 (deposit number FERM P-18932).

  Cellulase preparation Cellulase A “Amano” 3 and cellulase preparation Cellulosin AC40 (HB eye) 5 g each and protease preparation XP for 1 liter of citric acid added to the main culture solution to adjust the pH to 5 An enzyme treatment was performed by adding 2 g of -451 (Nagase Chemtex) and stirring at 50 ° C. for 8 hours. After completion of the enzyme treatment, pressure filtration through celite was performed to obtain a low molecular weight β-glucan aqueous solution.

Example 3 Production Example 3 of Microbe-Derived Low Molecular Weight β-Glucan
After culturing in the same manner as in Example 2, the cells were separated by centrifugation (5000 rpm, 20 minutes).

  Add 1g of glucanase preparation Cellulase Nagase (Nagase Chemtex) and 5g of Cellulase preparation Cellulase Y-NC (Yakult) to 1L of citric acid added to the main culture filtrate and adjust pH to 4. Pressure homogenizer (LAB1000 type) , APV), the homogenization treatment at 50 MPa was repeated twice, and then the mixture was stirred at 50 ° C. for 6 hours to carry out the enzyme treatment. Thereafter, the pH was adjusted to 7 with sodium hydroxide to obtain a low molecular weight β-glucan aqueous solution.

Example 4 Production Example 4 of Microbial Derived β-Glucan
In Example 2, 15 g of a pectinase preparation pectinase G “Amano” sterilized by filtration when 24 hours passed after inoculation of the seed culture solution was added, and the cellulase preparation cellulase Y-NC ( Yakult) 20 g was added, cultured for 24 hours, and finally cultured for 72 hours. Culture was performed in the same manner as in Example 2 and modified β-glucan was prepared at the same time as the culture was completed. Filtration was performed in the same manner as in Example 2 to obtain a low molecular weight β-glucan aqueous solution.

Example 5 Production Example 5 of Microbe-Derived Low Molecular Weight β-Glucan
Bacteria were separated from the culture solution obtained by the same method as in Example 1 by centrifugation, and 4 L of ethanol was added to 2 L of culture diluted solution to which an equal amount of water was added to the culture solution to obtain a precipitate. Only the precipitate was separated and dissolved in 1 L of water, and then 2 L of ethanol was added again to precipitate again. After performing the same operation on the precipitate again, the obtained precipitate was dissolved in 1.5 L of water. This was freeze-dried to obtain a freeze-dried product of β-glucan.

  The obtained dried glucan was dissolved in 500 mL of water and adjusted to pH 5 with lactic acid, and then 5 g each of cellulase preparation Cellulosin AC40 (HB i) and glucanase preparation Cellulase Nagase (Nagase Chemtex) were added. , And stirred at 40 ° C. for 16 hours to obtain a low molecular weight β-glucan.

Example 6 Production Example 6 of Microbe-Derived Low Molecular Weight β-Glucan
After culturing in the same manner as in Example 2, the cells were separated by centrifugation (5000 rpm, 20 minutes).

  Citric acid was added to 1 L of the main culture filtrate so that the citric acid concentration was 5%, heated at 100 ° C. for 4 hours, adjusted to pH 5 with sodium hydroxide, and 8 g of glucanase preparation Cellulase Nagase (Nagase Chemtex) And 5 g of cellulase preparation cellulase Y-NC (Yakult) was added and stirred at 50 ° C. for 3 hours for enzyme treatment. 2 L of water was added to this solution, followed by 3 L of ethanol, and the precipitate was recovered. The precipitate was dissolved in 1 L of water, 2 L of ethanol was added again, and the resulting precipitate was air-dried and then crushed to reduce the molecular weight. β-glucan powder was obtained.

Comparative Example 1
The same operation as in Example 1 was carried out except that the enzyme was added and the operation of stirring at 40 ° C. for 20 hours was not carried out to obtain a β-glucan solution.

Comparative Example 2
The same operation as in Example 2 was carried out except that the enzyme was added and the operation of stirring at 50 ° C. for 8 hours was not performed to obtain a β-glucan solution.

Comparative Example 3
The same operation as in Example 3 was carried out except that the enzyme was added and the operation of stirring at 50 ° C. for 6 hours was not performed to obtain a β-glucan solution.

Comparative Example 4
The same operation as in Example 5 was carried out except that the enzyme was added and the operation of stirring at 40 ° C. for 16 hours was not carried out to obtain a β-glucan solution.

Comparative Example 5
The same operation as in Example 6 was carried out except that the enzyme was added and the operation of stirring at 50 ° C. for 3 hours was not carried out to obtain β-glucan powder.

Analysis Example 1 (Method for confirming β-glucan and method for measuring β-glucan content)
For analysis of β-glucan in polysaccharide, the total amount of polysaccharide precipitated by alcohol is measured by the phenol-sulfuric acid method, and then β-glucan in the precipitated polysaccharide is confirmed and quantified by β-1 of Seikagaku Corporation. This was carried out using a β-glucan detection / measurement kit containing a 3-D-glucopyranose bond. The procedure will be described below.

  First, the total polysaccharide amount in the measurement sample was measured by the phenol sulfuric acid method. That is, 30 μL of distilled water was added to 30 μL of the sample solution, 120 μL of phosphate buffer solution (pH 6.9) containing 300 mM of NaCl was added thereto, 540 μL of ethanol (3 times volume) was added, and the mixture was added to −15 ° C. for 10 minutes. The polysaccharide was left to stand. After removing the supernatant, 100 μL of distilled water was added to dissolve the precipitate. After 100 μL of 5 wt% phenol aqueous solution and 500 μL sulfuric acid were added and reacted, the absorbance at 490 nm of the solution was measured. Further, the absorbance at 490 nm of the blank was measured using a blank obtained by adding 100 μL of a 5 wt% phenol aqueous solution and 500 μL of sulfuric acid to 100 μL of distilled water without adding a sample. Create a 2-fold dilution series from pullulan 10 mg / mL to make a standard sample, create a calibration curve using the standard sample, and quantitate the total amount of polysaccharides in the sample from the calibration curve and the absorbance above did.

Next, a solution having a total polysaccharide content of about 0.1 to 1 mg / mL is first diluted 10-fold with 1.0 M NaOH, then diluted with β-glucan-free distilled water, and diluted to 10 ^10- fold. A β-glucan diluted solution 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 then the absorbance of the solution at 545 nm (control wavelength: 630 nm) was measured. In addition, a β-60 glucan solution of 7.5-60 pg / mL was prepared using the attached β glucan standard, and a calibration curve was obtained. From the calibration curve and the absorbance, the concentration of each β-glucan diluted solution was calculated to determine the amount of β-glucan in the sample.

  As a result, the contents of low molecular β-glucan in Examples 1 to 5 were 0.72, 1.01, 0.93, 0.96, and 1.78%, respectively, and β-glucan in Comparative Examples 1 to 4 The contents of were 0.77, 1.05, 0.98, and 1.80%, respectively.

Evaluation Example 1 (Manufacturability during production)
The time required for Celite filtration in Example 2 and Comparative Example 2 was 10 minutes and 40 minutes, respectively, and it was found that the manufacturing process can be greatly shortened by the modification effect of the present invention.

Evaluation Example 2 (Handling property during production)
The β-glucan obtained in Examples 1 to 5 and Comparative Examples 1 to 4 was spray-dried (Labulus-1 type manufactured by Ashizawa Spray Plant). The β-glucan obtained in each example gave a clean spray-dried product, and the yield was 90 to 93%. However, the β-glucan obtained in the comparative example was mixed with a cocoon-like dried product by stringing during spraying. In addition, there was much adhesion of the undried product to the wall, and the yield was 49 to 53%. Therefore, it became clear that the handling property at the time of manufacture becomes very good.

Evaluation Example 3 (Handling property during production)
The β-glucan obtained in Examples 1 to 5 and Comparative Examples 1 to 4 was concentrated with a rotary evaporator to prepare a concentrated solution. As a result, as the concentration ratio without losing fluidity, the examples were 6, 8, 9, 5, 10 times in order from 1, whereas in the comparative examples, 3, 3, 4, 3 times in order from 1. there were. Therefore, it became clear that the handling property at the time of manufacture becomes very good.

Evaluation Example 4 (Breadmaking test)
A bread baking test was conducted with the goal of blending 50 mg of β-glucan per 6 pieces of cut bread (60 g). As the bread baking results vary to some extent, a recipe is added in which 333 mg (50/60 × 400) of β-glucan is added to a recipe that averages 400 g of 1 kg after baking. A bread making test was conducted.

  Formulation is 254 g of flour, 7.6 g of yeast, 10 g of sugar, 5 g of salt, 10 g of shortening, 160 g of β-glucan and water obtained in Examples 1 to 5 and Comparative Examples 1 to 4, and among them, Analysis Example 1 The addition amount of β-glucan aqueous solution calculated from the results of Examples 1-5 is 46.3, 33.0, 35.8, 34.7, 18.7 g in order, and Comparative Examples 1-4 are in order. 43.2, 31.7, 34.0, 18.5 g. These calibrations were added, and the dough was prepared by mixing with a hopper mixer at a low speed of 2 minutes and at a high speed of 4 minutes at a kneading temperature of 28 ° C. After fermentation at 28 ° C. for 60 minutes, the product was rounded, crushed (28 ° C., 20 minutes), passed through a sheeter three times, shaped, and then inserted into a one-loaf type mold. A roasting process was performed until the dough reached 2 cm at the upper edge of the mold at 38 ° C. and a relative humidity of 90%, followed by baking at 220 ° C. for 23 minutes.

  In addition to evaluating the uniformity of the dough during mixing at the time of bread dough creation and handling at the time of bread making from adhering to the blades of the mixer, the inner phase of the bread after baking, flavor, melting in the mouth, elasticity, Evaluation by 5 panelists was performed with a maximum score of 10 points, and the average score is shown in Table 1. As a result, in general, the β-glucan not treated with the enzyme of the comparative example has a non-uniform dough at the time of mixing, and as a result, the state of the inner phase of the baked bread is bad, and the taste and texture are remarkably inferior. became.

Evaluation Example 5 (Creation of tea beverage)
The powdered β-glucan obtained in Example 6 and Comparative Example 5 and the powdered β-glucan derived from Example 3 and Comparative Example 3 obtained in Evaluation Example 2 are each 2% by weight in a commercially available tea beverage (lemon tea). The panel was evaluated by five panelists for the flavor and throat, and the average score was shown in Table 2. As a result, the β-glucan that had not been subjected to the enzyme treatment did not deteriorate the flavor so much, but a smooth throat was not obtained, and the texture was not impaired at all by the enzyme treatment.

Example 7 Production Example of Barley-Derived β-Glucan Sticky bare barley was shaved with a grinding mill and refined to a yield of 82%. The soot generated at this time was designated as 糠 -1. The barley that had been milled to a yield of 82% was further shaved by a grinding mill, and was milled to a yield of 55%. The soot generated at this time was designated as pulverized product-1. 20 L of tap water was added to the container (50 L), and the temperature was adjusted to 15 ° C. while stirring. 6 kg of ６-1 was added thereto, and the mixture was extracted by stirring for 2 hours. After solid-liquid separation with a continuous centrifuge, the supernatant was lyophilized to obtain 450 g of an extraction accelerator. To the container (70 L), 30 L of tap water was added, and 150 g of the extraction accelerator was added while stirring, and after dissolution, 7.5 kg of the pulverized product-1 was added. After stirring and extracting at 50 ° C. for 2 hours, the supernatant was obtained after solid-liquid separation with a continuous centrifuge. The obtained supernatant was boiled, and after cooling, 15 L of a slightly viscous β-glucan solution was obtained. Two times the amount of ethanol was added to the obtained β-glucan solution, and the precipitate was collected and dried to obtain 460 g of β-glucan extracted from barley (sample A). Sample A had a weight average molecular weight of 100,000.

Example 8 Production Example of Microbe-Derived (Microbial Extracellular Production) β-Glucan The ADK-34 strain, which is an Aureobasidium pullulans strain having the deposit number FERM BP-8391, is cultured in a potato dextrose agar slant medium. The strain was inoculated into a 500 ml Erlenmeyer flask containing 100 ml of YM liquid medium (Difco) and pre-cultured at 28 ° C. for 3 days. In the main culture, 15 liters of Czapeak's medium (manufactured by Difco) and the resulting preculture were added to a 30-liter fermenter equipped with full zone blades and cultured at 28 ° C. for 3 days. During the culture, the pH was adjusted to 5.0, and the aeration amount and the number of rotations were controlled in sequence so that the aeration was 1 vvm. After 15 liters of the culture broth was sterilized by heating at 90 ° C. for 30 minutes, an equal amount of water was added, and then the cells were removed by centrifugation. The resulting culture supernatant was directly used as a UF membrane separator (UFP-10C-). 8A, manufactured by Amersham Bioscience) and desalted. The desalted culture supernatant was freeze-dried to obtain 110 g of a β-glucan freeze-dried product (Aureobasidium culture: sample B). Sample B had a weight average molecular weight of 2 million.

Example 9 Production Example of Basidiomycete-derived β-glucan Fruit body of Kawariharatake was crushed and pulverized, 50 liters of hot water was added to 10 kg of the pulverized product, and hot water extraction was performed for 3 hours with gentle stirring under boiling conditions. Processed. After the hot water extraction treatment, the solution was centrifuged to obtain the separated solution. Three times the amount of 99% ethyl alcohol was added to the separated liquid to obtain a precipitate, which was freeze-dried to obtain 1200 g of β-glucan (sample C). Sample C had a weight average molecular weight of 1,000,000.

Example 10 β-glucan complex 50 parts of barley-derived β-glucan (Sample A) obtained in Example 7 and 50 parts of the microorganism-derived β-glucan (Sample B) obtained in Example 8 were mixed to obtain β-glucan. A composite (sample D) was obtained.

Example 11 Production Example of Margarine 100 parts of edible oil and fat containing palm oil: palm hardened oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex (sample D) obtained in Example 10, and the mixture was allowed to stand at 65 ° C. for 30 minutes, and then stirred with a homomixer and heated to 70 ° C. with water 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 12 β-glucan complex 50 parts of barley-derived water-soluble β-glucan (Sample A) obtained in Example 7 and 50 parts of basidiomycete-derived β-glucan (Sample C) obtained in Example 9 were mixed, A β-glucan complex (sample E) was obtained.

Example 13 Production Example of Margarine 100 parts of edible oil and fat containing palm oil: palm hardened oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex (sample E) obtained in Example 12, and the mixture was allowed to stand at 65 ° C. for 30 minutes, and then stirred with a homomixer and water 16 heated to 70 ° C. 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 14 β-glucan complex 50 parts of the microorganism-derived β-glucan (sample B) obtained in Example 8 and 50 parts of the basidiomycete-derived β-glucan (sample C) obtained in Example 9 were mixed, and β A glucan complex (sample F) was obtained.

Example 15 Production Example of Margarine 100 parts of edible fat and oil containing palm oil: palm hardened oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex (sample F) obtained in Example 14, and the mixture was allowed to stand at 65 ° C. for 30 minutes, and then stirred with a homomixer and heated to 70 ° C. with water 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 16 β-glucan complex 40 parts of barley-derived water-soluble β-glucan obtained in Example 7 (sample A), 30 parts of the microorganism-derived β-glucan (sample B) obtained in Example 8 and basidiomycetous-derived β Glucan (sample C) was mixed to obtain a β-glucan complex (sample G).

Example 17 Production Example of Margarine 100 parts of edible oil and fat containing palm oil: hardened palm oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex (sample G) obtained in Example 16, and the mixture was allowed to stand at 65 ° C. for 30 minutes, and then stirred with a homomixer and heated to 70 ° C. with water 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 18 Production Example of Barley-Derived Low-Molecular (Water-Soluble) β-Glucan Sticky bare barley was shaved with a grinding mill and refined to a yield of 82%. The soot generated at this time was designated as 糠 -1. The barley that had been milled to a yield of 82% was further shaved by a grinding mill, and was milled to a yield of 55%. The soot generated at this time was designated as pulverized product-1. Add 30 L of tap water to a container (70 L), add 7.5 kg of the pulverized product-1 while stirring, add 300 g of cellulase preparation cellulase A “Amano” 3 (Amanoenzyme), and continue at 50 ° C. for 2 hours. After stirring and extraction, the supernatant was obtained after solid-liquid separation with a continuous centrifuge. The obtained supernatant was boiled, and after cooling, 15 L of a slightly viscous β-glucan solution was obtained. Two times the amount of ethanol was added to the obtained β-glucan solution, and the precipitate was collected and dried to obtain 600 g of low molecular weight (water-soluble) β-glucan derived from barley.

Example 19 Production Example of Margarine 100 parts of edible oil and fat containing palm oil: hardened palm oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of a low molecular weight (water-soluble) β-glucan extracted from the barley obtained in Example 18, and left at 65 ° C. for 30 minutes, and then stirred at 70 ° C. with a homomixer. After gradually adding and mixing a solution in which 0.5 part of skim milk powder and 1 part of sodium chloride was dissolved in 16 parts of warm water, quench plasticization was performed, and the temperature was adjusted overnight at 25 ° C. until 5 ° C. Cooled to obtain margarine. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 20 β-glucan complex 50 parts of the low-molecular-weight β-glucan powder derived from microorganisms obtained in Example 6 and 50 parts of the low-molecular-weight (water-soluble) β-glucan derived from barley obtained in Example 18 were mixed. Β-glucan complex (sample H) was obtained.

Example 21 Production Example of Margarine 100 parts of edible fat and oil containing palm oil: palm hardened oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex (sample H) obtained in Example 20, and the mixture was allowed to stand at 65 ° C. for 30 minutes, and then stirred with a homomixer and heated to 70 ° C. with water 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 22 Production Example of Low Molecular Weight β-Glucan Derived from Basidiomycetes The fruit body of Kawariharatake is crushed and pulverized, 50 liters of water is added to 10 kg of the pulverized product, and 400 g of cellulase preparation cellulase A “Amano” 3 (Amanoenzyme) is used. After adding and stirring and extracting at 50 ° C. for 2 hours, solid-liquid separation was performed with a continuous centrifuge, and the separated liquid was obtained. Three times the amount of 99% ethyl alcohol was added to the separated liquid to obtain a precipitate, which was lyophilized to obtain 1300 g of a low molecular weight β-glucan derived from basidiomycetes.

Example 23 β-glucan complex 50 parts of a low molecular weight β-glucan derived from basidiomycete obtained in Example 22 and 50 parts of a low molecular weight (water-soluble) β-glucan derived from barley obtained in Example 18 were mixed. A β-glucan complex (Sample I) was obtained.

Example 24 Production Example of Margarine 100 parts of edible oil and fat containing palm oil: hardened palm oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex obtained in Example 23 (Sample I), left at 65 ° C. for 30 minutes, and then stirred with a homomixer and heated to 70 ° C. with water 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

Example 25 β-glucan complex 50 parts of the microorganism-derived β-glucan (sample B) obtained in Example 8 and 50 parts of a low molecular weight (water-soluble) β-glucan derived from barley obtained in Example 18 were mixed. A β-glucan complex (Sample J) was obtained.

Example 26 Production Example of Margarine 100 parts of edible oil and fat containing palm oil: palm hardened oil: rapeseed oil: sorbitan fatty acid ester in a ratio (weight ratio) of 30: 50: 20: 0.3 was melted at 70 ° C. To this was added 8 parts of the β-glucan complex (sample J) obtained in Example 25, and the mixture was allowed to stand at 65 ° C. for 30 minutes, and then stirred with a homomixer and heated to 70 ° C. with water 16 After gradually adding and mixing a solution in which 0.5 parts of skim milk powder and 1 part of sodium chloride are mixed, quench plasticization is performed, and the mixture is conditioned overnight at 25 ° C and then cooled to 5 ° C to obtain margarine. It was. The β-glucan complex was uniformly dispersed. The obtained margarine was excellent in flavor and had a fine and smooth texture.

  In the following examples, the molecular weight and β-glucan content of β-glucan were measured according to the following analysis examples 2 and 3, respectively. In the following examples, the molecular weight indicates a weight average molecular weight.

Analysis Example 2 (Method for measuring molecular weight of β-glucan)
The molecular weight of β-glucan was measured as follows.
That is, 5 mg of β-glucan was taken in a tube, 1 ml of distilled water was added, and dissolved in boiling water. A sample for HPLC was obtained through a 0.45 μm filter. For separation, Shodex packed column KS-804 (manufactured by Showa Denko), which is an HPLC gel filtration column, was used at a flow rate of 0.6 ml / min. At a temperature of 80 ° C., detection was performed using an RI detector, and water was used as a separation solvent. The molecular weight marker was measured using Shodex pullulan standard solutions P-5 to P160 (manufactured by Showa Denko).

Analysis Example 3 (Method for measuring β-glucan content)
The β-glucan content was measured by the McCleary method (enzyme method) as follows using a β-glucan measurement kit manufactured by Megazyme.
First, the water content of each of the fractions passed through a 500 μm (30 mesh) sieve was measured, 10 mg of the fraction 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 acetate buffer, and the remaining amount was stored frozen) and reacted at 50 ° C. for 10 minutes. 3 ml of glucose oxidase / peroxidase 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 (%, w / w) = (EA) × (F / W) × 8.46
F = (100) / (absorbance of glucose 100 μg)
W = calculated anhydride weight (mg)

Example 27 (Preparation of enzyme-treated product)
First, β-glucan was extracted according to JP-A-2002-105103.
That is, glutinous naked barley was shaved with a grinding mill and polished to 10% (milling yield 90%). 200 g of the pulverized soot generated at this time was placed in a beaker, 1 L of distilled water was added, and the mixture was stirred and extracted at 25 ° C for 10 minutes. After extraction, the supernatant obtained by centrifugation and filtration was designated as Extract-1.

  Commercially available barley grains (50% yield) were crushed into barley flour. 8 L of tap water was added to 2 kg of barley flour, 800 ml of Extract 1 was added with stirring, and the mixture was extracted by stirring at 55 ° C. for 2 hours. After extraction, the mixture was centrifuged to obtain a supernatant, and 1 L of a concentrated solution was obtained using an evaporator. An equal amount of ethanol was added, and the mixture was allowed to stand for 1 hour, and then the precipitate was collected by centrifugation. Ethanol / water was removed with a freeze dryer to obtain β-glucan extract-1 (molecular weight 60,000, β-glucan content 66%) from barley. Using this as a raw material, an enzyme was added, and a low molecular weight β-glucan was prepared as follows. The component composition of β-glucan extract-1 is shown in Table 3.

  Add 380 ml of water to 20 g of β-glucan extract-1 and dissolve at a temperature of 60 ° C. While maintaining the temperature at 50 ° C., add 20 ml of enzyme solution (Ceremix, 0.1% aqueous solution) and mix well. Thereafter, 20 ml of the reaction solution was collected over time (sampling time was 0.1 to 24 hours), and the enzyme was inactivated by treatment at 121 ° C. for 20 minutes. Each reaction solution was freeze-dried to obtain an enzyme-treated product. The molecular weight of the enzyme-treated product obtained by sampling over time was measured according to Analysis Example 2. Sample K0 (molecular weight 60,000), which is a raw material, and each enzyme-treated product which is the water-soluble β-glucan of the present invention, ie, sample K1 (molecular weight 30,000), sample K2 (molecular weight 15,000), sample K3 (molecular weight) 8000), sample K4 (molecular weight 6000), sample K5 (molecular weight 4000), sample K6 (molecular weight 2000) and sample K7 (molecular weight 800) were used in Test Example 1 below. Celexix is a liquid protease preparation enzyme in which α-amylase, β-glucanase and protease are mixed, and is an enzyme produced by Bacillus amyloliquefaciens.

Example 28 (Preparation of enzyme-treated product)
Except that barley extract-2 (molecular weight: 40,000, β-glucan content: 74%) extracted from barley was used, and the enzyme for lowering the molecular weight was replaced with Grainase (manufactured by Daiwa Kasei Kogyo Co., Ltd.). Thus, an enzyme-treated product was obtained. Sample L0 (molecular weight 40,000) as a raw material, and each enzyme-treated product that is the water-soluble β-glucan of the present invention, ie, sample L1 (molecular weight 20,000), sample L2 (molecular weight 10,000), sample L3 (molecular weight 7000) Sample L4 (molecular weight 5000), sample L5 (molecular weight 3000), sample L6 (molecular weight 2000) and sample L7 (molecular weight 1000) were used in Test Example 2 below. In addition, Glenase is a β-glucanase preparation and an enzyme produced by Bacillus subtilis.

Example 29 (Preparation of enzyme-treated product)
100 g of water was added to 1 g of β-glucan purified from oat (manufactured by Nippon Biocon, 97% purity) and dissolved at 60 ° C., and 5 μl of celeclast (manufactured by Novozyme) was added. After reacting for 5 hours, it was freeze-dried to obtain a powder (sample M) of the enzyme-treated product which is the water-soluble β-glucan of the present invention. Seleclast is a β-glucanase preparation and an enzyme produced by Trichoderma reesei.

Example 30 (Preparation of enzyme-treated product)
100 g of water is added to 1 g of β-glucan (manufactured by Nippon Biocon, low viscosity product) purified from barley, dissolved at 60 ° C., 1 μl of Grainase (manufactured by Daiwa Kasei) is added, and then at 30 ° C. for 24 hours. After the reaction, freeze-dried to obtain an enzyme-treated product powder (sample N) which is the water-soluble β-glucan of the present invention.

Example 31
Commercial barley was pulverized into barley flour. The β-glucan content of the barley flour was 4.6%. Five times the amount of water was added to 100 g of barley flour, gently stirred at 40 ° C. for 4 hours, and a supernatant was obtained by centrifugation. This solution was concentrated to 200 ml with an evaporator, slowly cooled to −30 ° C., frozen, thawed after 2 days, and the precipitate was collected and lyophilized to obtain 3.2 g of barley extract-3. Barley extract-3 was dissolved again in 500 ml of distilled water, slowly cooled to −30 ° C., frozen, thawed after 2 days, and the precipitate was collected and lyophilized to obtain barley extract-4. 1 g was obtained. The same operation was performed again to recover the precipitate, and half of the precipitate was suspended in 20 ml of distilled water to obtain barley extract-1. The other half was freeze-dried to obtain 0.8 g of barley extract-5. When β-glucan content and molecular weight were measured according to the analysis example, barley extract-5 had a β-glucan content of 89% and a molecular weight of 130,000. After adding 10 U of lichenase (manufactured by Nippon Biocon) to 100 mg of this barley extract-5 and reacting for 5 hours, the enzyme was inactivated by heat treatment at 90 ° C. for 2 hours to obtain the water-soluble property of the present invention. An enzyme-treated product (sample O) that was β-glucan was obtained. Moreover, after adding lichenase 2U to 1 ml of barley extract-1 and carrying out an enzyme reaction until precipitation lose | disappears at 50 degreeC, an enzyme is deactivated by 90 degreeC and the heat processing for 2 hours, An enzyme-treated product (sample P) that was a water-soluble β-glucan was obtained.
The lichenase is an enzyme produced by Bacillus subtilis and is endo-β-D-glucanase (EC 3.2.1.73).

Example 32
30 ml of the enzyme solution was added to 10 g of barley extract-6 (molecular weight 80,000) having a β-glucan content of 80%, and the mixture was well stirred and mixed in a mortar and left at 50 ° C. for 2 hours. After completion of the reaction, the enzyme was inactivated at 90 ° C. for 3 hours, and then freeze-dried to obtain an enzyme-treated product (sample Q) that is the water-soluble β-glucan of the present invention. As the enzyme solution, a 0.01% aqueous solution of Grainase (manufactured by Daiwa Kasei Co., Ltd.) was used.

Example 33
Add 50 ml of distilled water to 50 g of commercially available oat flakes (Quaker Oatmeal: made by Snow Brand), stir for 1 hour, slowly cool to −30 ° C., freeze, thaw spontaneously after 2 days, centrifuge and precipitate Was recovered. 80 ml of distilled water was added to 10 g of the freeze-dried product, 5 U of lichenase (manufactured by Nippon Biocon) was added with stirring, and the mixture was reacted at 40 ° C. for 3 hours, and then at 90 ° C. for 1 hour. The enzyme was inactivated and freeze-dried to obtain an enzyme-treated product (sample R) which is the water-soluble β-glucan of the present invention.

Example 34
3 g of barley extract-6 was added to 100 ml of 0.5% enzyme preparation aqueous solution (Celemix: Novozyme), reacted at 50 ° C. with stirring, and further 2 hours, 4 hours and 6 hours later 8 hours later, 3 g of barley extract-6 was added and allowed to react for 24 hours. Subsequently, the enzyme was inactivated at 90 ° C. for 2 hours and then freeze-dried to obtain an enzyme-treated product (sample S) which is the water-soluble β-glucan of the present invention.

Test Example 1 (Solubility test)
Test sample:
Commercially available β-glucan and enzyme-treated products of β-glucan obtained in Examples 27 and 28 (samples K0 to K7, L0 to L7) were used as samples to be tested. Commercially available β-glucan having a molecular weight of 240,000 (sample a) and 125,000 (sample b) was used (manufactured by Nippon Biocon).
Test method:
Each sample was added to distilled water adjusted to 25 ° C. so as to have a concentration of 20 mg / ml, and dissolved by stirring for 10 minutes at room temperature. This solution was left at 4 ° C. for 3 days. The presence or absence of precipitation in the solution was visually confirmed at 3 points after dissolution at room temperature and after 1 day from the start of standing at 4 ° C. and after 3 days. The results are shown in Table 4. In the evaluation of solubility, precipitation was observed without dissolution (x), hydration and dispersion were achieved, precipitation was not observed (◯), and dissolution was observed without precipitation (◎). As is apparent from the results in Table 4, the solubility increased with a decrease in the molecular weight, complete dissolution was achieved at a molecular weight of 10,000 or less, and no precipitation was observed at a molecular weight of 20,000 or less during storage at 4 ° C.

Test Example 2 (Solubility test)
Samples to be tested:
Commercially available β-glucan and the enzyme-treated product of β-glucan obtained in Example 27 (samples K0 to K7) were used as samples to be tested. Commercially available β-glucan having a molecular weight of 240,000 (sample a) and 125,000 (sample b) was used (manufactured by Nippon Biocon).
Test method:
Each sample was added to distilled water to a concentration of 50 mg / ml and completely dissolved at 60 ° C. The sample solution was allowed to stand at room temperature, 4 ° C. or −30 ° C. for a whole day and night, and then the sample was brought to room temperature to observe the properties and measure the absorbance of the solution at 660 nm. In the property observation, the presence or absence of gelation and the viscosity are observed. The gel state is completely gelled (+++), the gel state is crumbly broken (++), the liquid state is viscous without gelation (+), the viscosity is The state of each sample was evaluated as a low liquid state (±). Absorbance measurement at 660 nm was carried out with a 96-well microplate reader (Bio-Rad) using 200 μl of the well-mixed solution at room temperature. The sample with the highest absorbance value was taken as 100, and the turbidity of each sample was shown as a relative value. The results are shown in Table 5.

  As is clear from Table 5, the properties show the same tendency for both storage at room temperature and storage at 4 ° C., and solid gelation is observed in samples a and b having molecular weights of 245,000 and 120,000. The solution was solidified. When the molecular weight was 60,000, it was in a soft gel state, and when the molecular weight was 30,000, it was a viscous liquid. When the molecular weight was 10,000 or less, no gelation was observed and the solution was in a solution state. Moreover, in the sample preserve | saved at -30 degreeC, even after returning to room temperature, strong gelling is recognized by the sample a and b of molecular weight 240,000 and 1280,000, and it becomes a soft gel state by molecular weight 60,000. The sample had a molecular weight of 30,000 and was a viscous liquid. A sample having a molecular weight of 6000 or less became a non-viscous liquid.

  Further, as a result of measuring the absorbance, as is apparent from Table 5, among samples stored at room temperature and 4 ° C., the sample having a molecular weight of 60,000 has the highest absorbance of 0.3. It was 50 or less for samples having a molecular weight of 8000 or less, and it was almost transparent and no turbidity was observed by visual observation. Samples a and b were completely gelled and could not be measured. Moreover, in the sample preserve | saved at -30 degreeC, the light absorbency of the sample with a molecular weight of 60,000 is the highest with 1.5, and also the light absorbency falls with the fall of a molecular weight. When the molecular weight was 6000 or less, the relative absorbance was 1 or less, and it was almost transparent and no turbidity was observed by visual observation.

Test Example 3 (Solubility test)
Samples to be tested:
Commercially available β-glucan and the enzyme-treated product of β-glucan obtained in Example 28 (samples L0 to L7) were used as samples to be tested. As the commercially available β-glucan, barley β-glucan (manufactured by Nippon Biocon) having a molecular weight of 240,000 (sample a) and 125,000 (sample b) was used as a control.
Test method:
Each sample was added to distilled water to a concentration of 10 mg / ml and dissolved at 60 ° C. The sample solution was allowed to stand at −30 ° C. for a whole day and night, then the sample was returned to room temperature, and property observation and the absorbance at 660 nm of the solution were measured. Evaluation was the same as in Test Example 2. That is, in the property observation, the presence or absence of gelation and the viscosity are observed, and the gel is completely gelled (+++), the gel is crumbly broken (++), or the liquid is viscous without being gelled (+) The state of each sample was evaluated as a liquid state (±) with low viscosity. The absorbance at 660 nm was measured by a 96-well microplate reader (manufactured by Bio-Rad) using 200 μl of the solution which had been returned to room temperature. The sample with the highest absorbance value was taken as 100, and the turbidity of each sample was shown as a relative value. The results are shown in Table 6.

As is clear from Table 6, after leaving at −30 ° C., a sample with a molecular weight of 40,000 to 10,000 became a viscous liquid, and a sample with a molecular weight of 7,000 or less became a low viscosity liquid.
Further, as a result of measuring the absorbance, the absorbance of the sample having a molecular weight of 40,000 is the highest, 0.801, and the absorbance decreases with a decrease in the molecular weight. If the absorbance of the sample having the molecular weight of 40,000 is defined as the relative absorbance of 100, the molecular weight is 10,000 or less. In Comparative Example 1, the relative absorbance was 10 or less, and it was almost transparent and no turbidity was observed by visual observation.

Test example 4
The β-glucan content and molecular weight in the samples obtained in Examples 29 to 34 were examined.
In addition, each sample was dissolved in distilled water so as to be 30 mg / ml and stored at 4 ° C. all day and night, and then the state of gelation and precipitation was observed. As a control, the sample a and the sample b were evaluated together. In the property observation, the presence or absence of gelation and the viscosity are observed, and the gel is completely gelled (+++), the gel is crumbly broken (++), the liquid is viscous without being gelled (+), or the liquid is low in viscosity The state of each sample was evaluated as the state (±). Sample R was evaluated by adding 10 times the amount of distilled water and extracting for 10 minutes, and then lyophilizing the supernatant obtained by centrifugation as sample R2. The results are shown in Table 7. All the enzyme-treated products were found to have good solubility and excellent storage stability.

Claims (18)

  Β-glucan derived from microorganisms.   The β-glucan derived from microorganisms according to claim 1, which is β1-3, 1-6 glucan.   Cereal-derived β-glucan.   The β-glucan derived from cereal according to claim 3, which is β1-3, 1-4 glucan.   Β-glucan derived from basidiomycetes.   The β-glucan derived from basidiomycetes according to claim 5, which is β1-3 or 1-6 glucan.   A β-glucan complex containing β1-3, 1-6 glucan and β1-3, 1-4 glucan.   A β-glucan complex containing at least two of β-glucan derived from microorganisms, β-glucan derived from grains, and β-glucan derived from basidiomycetes.   Β-glucan complex containing any one or more of microbial-derived β1-3,1-6 glucan, cereal-derived β1-3,1-4 glucan, basidiomycetous-derived β1-3,1-6 glucan .   Water-soluble β-glucan obtained by adding an enzyme to cereal and / or cereal-derived β-glucan to reduce the molecular weight.   The water-soluble β-glucan according to claim 10, obtained by lowering the molecular weight by adding an enzyme to a grain containing hemicellulose and / or a grain-derived β-glucan containing hemicellulose.   The water-soluble β-glucan according to claim 10 or 11, which is obtained by reducing the molecular weight of barley or oat and / or β-glucan derived from barley or oat by adding an enzyme to act.   The water-soluble β-glucan according to any one of claims 10 to 12, wherein the cereal and / or the cereal-derived β-glucan is an extract or a fraction extracted from the cereal.   The water-soluble β-glucan according to any one of claims 10 to 13, which has been reduced in molecular weight to a weight average molecular weight of 60,000 or less.   The water-soluble β-glucan according to any one of claims 10 to 14, wherein the enzyme to be added and acted on is an enzyme that catalyzes a reaction of hydrolyzing between β-D-glucose bonds.   A method for producing a low molecular weight β-glucan derived from microorganisms, which comprises a step of allowing a hydrolase to act.   A method for producing a low-molecular-weight β-glucan derived from cereal having a step of allowing a hydrolase to act. A method for producing a low molecular weight β-glucan derived from a basidiomycete having a step of allowing a hydrolase to act.