Method for producing composition and method for producing food or beverage

Abstract

To provide a method for producing a composition containing equol and / or an equol derivative, which is resistant to browning even when heated, at least in its native state or in the presence of additional amino acids or the like.SOLUTION: The problem is solved by a method for producing a composition, comprising the following step (a), step (b1) and / or step (b2), and step (c): (a) a step of forming glucose and isoflavone aglycone from an isoflavone glycoside; (b1) a step of causing a microorganism capable of producing equol to produce equol from the isoflavone aglycone; (b2) a step of allowing a microorganism capable of producing an equol derivative to produce the equol derivative from the isoflavone aglycone; and (c) a step of decomposing the glucose so as to adjust the content of the glucose to be 0 g / L or more and less than 2.0 g / L in the composition, or 30 g / kg or less per solid content of the composition.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to a method for producing a composition, a method for producing a food or beverage, and a fermented composition. [Background technology]

[0002] Isoflavone glycosides contained in soybean hypocotyls and soybean hypocotyl extracts are converted to aglycones by β-glucosidase, and then further metabolized to equol and equol derivatives, etc. Because equol has a strong physiological effect similar to that of female hormones, its use has been proposed for preventing and improving menopausal symptoms and osteoporosis (Patent Document 1), preventing and treating skin aging and wrinkles (Patent Document 2), and alleviating allergic symptoms (Patent Document 3), etc. Equol derivatives, particularly 5-hydroxyequol, are known to have antioxidant effects (Non-Patent Document 1) and lifespan extension effects (Non-Patent Document 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication No. 2001-523258 [Patent Document 2] Special Publication No. 2002-511860 [Patent Document 3] Patent No. 4479505 [Non-patent literature]

[0004] [Non-Patent Document 1] Archives Biochem. Biophys. vol.356, pp133-141(1998) [Non-Patent Document 2] J. Chin. Pharm. Sci. vol.23,pp378-384(2014) Summary of the Invention [Problem to be solved by the invention]

[0005] The present inventors produced a composition containing equol and / or equol derivatives from isoflavone glycosides, and found that the composition browned when heated. In addition, when the composition was used as a raw material to produce food, and mixed with raw materials containing proteins, etc., and heated, the composition browned when heated. This was thought to be due to a reaction between reducing sugars and amino acids called the Maillard reaction. In addition, the Maillard reaction may have produced by-products such as acrylamide, which are said to be carcinogenic.

[0006] An objective of the present disclosure is to provide at least a method for producing a composition containing equol and / or an equol derivative that does not brown when heated, either as is or in the presence of additional amino acids or the like. [Means for solving the problem]

[0007] As a result of intensive research, the present inventors have found that when isoflavone glycosides are converted into aglycones, glucose, which is a reducing sugar, is generated, and since the glucose remains in the produced composition, the Maillard reaction proceeds by heating, causing the composition to brown. Furthermore, they have found that the above-mentioned problems can be solved by carrying out a process of decomposing the glucose contained in the composition. That is, the present disclosure includes at least the following:

[0008] [1] A method for producing a composition, comprising the following steps (a); (b1) and / or (b2); and (c). (a) producing glucose and isoflavone aglycone from isoflavone glycoside; (b1) allowing a microorganism capable of producing equol to produce equol from the isoflavone aglycone; (b2) allowing a microorganism capable of producing an equol derivative to produce the equol derivative from the isoflavone aglycone; and (c) decomposing the glucose so that the content of the glucose in the composition is 0 g / L or more and less than 2.0 g / L, or 30 g / kg or less per solid content of the composition; [2] The production method according to [1], wherein the step (c) is a step of decomposing the glucose by fermentation using a microorganism having sugar assimilation ability. [3] The method according to [2], wherein the microorganism capable of sugar assimilation is one or more microorganisms selected from the group consisting of butyric acid bacteria, lactic acid bacteria, and yeast. [4] The microorganism having the ability to utilize sugar is a microorganism belonging to the genus Clostridium. Organisms, microorganisms belonging to the genus Lactobacillus, microorganisms belonging to the genus Lactococcus, microorganisms belonging to the genus Leuconostoc, and One or more microorganisms selected from the group consisting of microorganisms belonging to the genus Saccharomyces. The method for producing a living organism according to [3]. [5] The production method according to [1], wherein the step (c) is a step of decomposing the glucose using an enzyme that decomposes glucose. [6] The method according to [5], wherein the enzyme that decomposes glucose is one or more enzymes selected from the group consisting of glucose oxidase and glucose dehydrogenase. [7] The production method according to any one of [1] to [6], wherein the step (b1) and / or the step (b2) and the step (c) are carried out simultaneously. [8] The method according to any one of [1] to [7], wherein the isoflavone glycoside is daidzin and the isoflavone aglycone is daidzein. [9] The method according to any one of [1] to [8], wherein the step (a) is a step of reacting an isoflavone glycoside with β-glucosidase to produce glucose and an isoflavone aglycone.

[10] A method for producing a food or beverage, comprising the following steps (a); (b1) and / or (b2); (c); and (d). (a) producing glucose and isoflavone aglycone from isoflavone glycoside; (b1) allowing a microorganism capable of producing equol to produce equol from the isoflavone aglycone; (b2) allowing a microorganism capable of producing an equol derivative to produce the equol derivative from the isoflavone aglycone; (c) decomposing the glucose so that the content of the glucose in the composition produced by the method including the steps (a), (b1) and / or (b2), and (c) is 0 g / L or more and less than 2.0 g / L, or 30 g / kg or less per solid content of the composition; and (d) a step of producing a food or beverage using a composition produced by a method including the steps (a), (b1) and / or (b2), and (c).

[11] A fermented composition comprising equol and / or an equol derivative and substantially free of glucose.

[12] The fermentation composition according to

[11] , wherein the substantially free of glucose means containing glucose in an amount of 0 g / L or more and less than 2.0 g / L.

[13] A composition comprising equol and / or an equol derivative and glucose, A fermentation composition, wherein the glucose content is 30 g / kg or less per solid content of the fermentation composition.

[14] The fermentation composition according to any one of

[11] to

[13] , which is used in a food or drink. Effect of the Invention

[0009] The present disclosure may have the effect of providing at least a method for producing a composition containing equol and / or an equol derivative, which has a reduced amount of glucose and does not undergo the Maillard reaction or brown when heated, either as is or in the presence of additional amino acids, etc. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Each configuration and their combinations in each embodiment are merely examples, and addition, omission, substitution, and other modifications of the configurations are possible as appropriate within the scope of the present disclosure. The present disclosure is not limited by the embodiments, but is limited only by the scope of the claims. In addition, each aspect disclosed in this specification can be combined with any other feature disclosed in this specification. In addition, a numerical range expressed using "~" means a range that includes the numerical values ​​before and after "~" as the lower and upper limits, and "A~B" means A or more and B or less.

[0011] In this specification, the accession numbers of strains beginning with the word DSM are numbers assigned to microorganisms stored at DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH). The accession numbers of the strains beginning with the word JCM are those of the Japan Collection of Microorganisms (JCM). The numbers given to microorganisms are stored in the RIKEN BioResource Research Center, Microbial Materials Division, 3-1-1 Takanodai, Tsukuba, Ibaraki Prefecture, Japan, 305-0074. and is available from the same institution. The accession number of the strain beginning with the word FERM is the number given to the microorganisms stored at the National Institute of Advanced Industrial Science and Technology (currently the National Institute of Technology and Evaluation Patent Organism Depositary Center, Postal Code: 292-0818, Address: Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture), and can be obtained from the institution. The accession numbers of the strains beginning with the words KCCM are those of the Korean Culture Collection (KCCM). This is the number given to microorganisms stored at the Kochi Center of Microorganisms (KCCM). It can be obtained from the agency.

[0012] As used herein, "microorganism" refers to eubacteria, archaea, or fungi, and does not include viruses, animals, or plants.

[0013] In this specification, decomposing glucose means inactivating the reducing end of glucose so that glucose does not undergo the Maillard reaction with amino acids, etc., and includes the oxidation of glucose and conversion to another compound by metabolism. The amino acid or the like is not particularly limited as long as it is a compound capable of undergoing a Maillard reaction with glucose, and examples of the compound that contains an amino group include amino acids, peptides, and proteins.

[0014] One embodiment of the present disclosure is a method for producing a composition, comprising the following steps (a); (b1) and / or (b2); and (c). (a) producing glucose and isoflavone aglycone from isoflavone glycoside; (b1) allowing a microorganism capable of producing equol to produce equol from the isoflavone aglycone; (b2) allowing a microorganism capable of producing an equol derivative to produce the equol derivative from the isoflavone aglycone; and (c) decomposing the glucose so that the content of the glucose in the composition is 0 g / L or more and less than 2.0 g / L, or 30 g / kg or less per solid content of the composition;

[0015] [(a) Process] The production method according to this embodiment includes the step of (a) producing glucose and isoflavone aglycone from an isoflavone glycoside.

[0016] Isoflavone glycosides refer to glycosides of isoflavones. Isoflavones are a type of polyphenol, a flavonoid whose basic structure is isoflavone. Isoflavones are found in large amounts in legumes such as soybeans, kudzu, red clover, and licorice. Examples of isoflavones in this embodiment include isoflavones derived from legumes containing isoflavones. Specifically, isoflavones derived from soybeans (sometimes referred to as "soybean isoflavones" or "soybean isoflavones" in the technical field and market, and are treated as synonymous in this disclosure), isoflavones derived from kudzu (sometimes referred to as "kudzu isoflavones" or "kudzu isoflavones" in the technical field and market, and are treated as synonymous in this disclosure), isoflavones derived from red clover (sometimes referred to as "red clover isoflavones" or "red clover isoflavones" in the technical field and market, and are treated as synonymous in this disclosure), and isoflavones derived from licorice (sometimes referred to as "licorice isoflavones" or "licorice isoflavones" in the technical field and market, and are treated as synonymous in this disclosure).

[0017] Examples of isoflavone glycosides include genistin, glycitin, daidzin, etc. In this embodiment, it may be a mixture of two or more. For example, it may be a mixture of daidzin, glycitin, and genistin. These are contained in soybean hypocotyl, soybean hypocotyl extract, etc., so for example, soybean hypocotyl extract can be used as a mixture of isoflavone glycosides.

[0018] Examples of isoflavone aglycones include daidzein, 6-hydroxydaidzein, dihydroxydaidzein, genistein, glycitein, biochanin A, formononetin, orobol, and coumestrol. In this embodiment, a mixture of two or more of them may be used. Note that the term "isoflavone aglycone" in this disclosure is sometimes referred to as "aglycone of isoflavones" in the technical field and market, and is treated as having the same meaning in this disclosure.

[0019] In this embodiment, preferably, the isoflavone glycoside is daidzin and the isoflavone aglycone is daidzein, or the isoflavone glycoside is genistin and the isoflavone aglycone is genistein. More preferably, the isoflavone glycoside is daidzin and the isoflavone aglycone is daidzein.

[0020] Methods for producing glucose and isoflavone aglycone from isoflavone glycosides include a method of reacting isoflavone glycosides with β-glucosidase and a method of culturing a microorganism that produces β-glucosidase in a medium containing isoflavone glycosides.

[0021] β-glucosidase is an enzyme that catalyzes the reaction of hydrolyzing β-glycosidic bonds of sugars. β-glucosidase is not limited as long as it has the above-mentioned enzyme activity, and examples of β-glucosidase include those derived from microorganisms belonging to the genera Aspergillus, Penicillium, Rhizopus, Pseudomonas, Pichia, and Lactococcus, those derived from higher plants, and those derived from animals. In addition, commercially available products such as Pectinase G (Amano Enzyme Co., Ltd.), an enzyme preparation containing β-glucosidase, can also be used.

[0022] In order to allow β-glucosidase to act on isoflavone glycosides, The amount of β-glucosidase added can be adjusted as appropriate, but is usually 10 U to 3000 U, preferably 20 U to 1000 U, and more preferably 50 U to 500 U, per 1 g of isoflavone glycoside.

[0023] The conditions for allowing β-glucosidase to act on isoflavone glycosides are not particularly limited as long as the β-glucosidase acts on the isoflavone glycosides and catalyzes the release of glucose, but the temperature is usually 15°C or higher and 65°C or lower, preferably 20°C or higher and 55°C or lower, and more preferably 30°C or higher and 45°C or lower. The pH is usually 2 or more and 9 or less, preferably 3 or more and 7 or less, and more preferably 4 or more and 6 or less.

[0024] The microorganism that produces β-glucosidase is not particularly limited, and one or more microorganisms can be used regardless of genus, species, or strain. For example, microorganisms belonging to the genera Aspergillus, Lactococcus, Streptococcus, Lactobacillus, Bifidobacterium, Penicillium, Pichia, Pseudomonas, and Rhizopus. Examples include: Preferred examples of the microorganisms include those belonging to the genera Aspergillus, Lactococcus, Streptococcus, Lactobacillus, and Bifidobacterium. More preferably, Aspergillus niger, Aspergillus Aspergillus oryzae, Aspergillus aculeatus, Lactococcus lactis, Streptococcus spp. Examples of microorganisms that may be included in the bacteria include those belonging to the genus Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium bifidum.

[0025] As the culture conditions for the microorganisms that produce β-glucosidase, culture conditions that are usually used in the culture of the above-mentioned microorganisms or culture conditions that are appropriately modified therefrom can be used.

[0026] Examples of the culture medium that can be used include BHI medium manufactured by Difco, ANAEROBE BASAL BROTH (ABB medium) manufactured by Oxoid, Wilkins-Chalgren Anaerobe Broth (CM0643) manufactured by Oxoid, GAM medium and modified GAM medium manufactured by Nissui Pharmaceutical Co., Ltd. In this specification, the term "medium" refers to a solution in which microorganisms can grow, including a minimal medium, and does not include a solution in which microorganisms cannot grow, such as water, a salt solution, or a buffer solution.

[0027] The medium is supplemented with an isoflavone glycoside, which can be added before or during the culture of the microorganism. The content of isoflavone glycosides is not particularly limited, but is usually from 0.5 g / L to 100 g / L, preferably from 1 g / L to 50 g / L, and more preferably from 2 g / L to 20 g / L.

[0028] Water-soluble organic matter can be added to the medium as a carbon source. Examples of water-soluble organic matter include the following compounds: sugars such as glucose, arabinose, sorbitol, fructose, mannose, sucrose, trehalose, and xylose; alcohols such as glycerol; organic acids such as valeric acid, butyric acid, propionic acid, acetic acid, formic acid, fumaric acid, and succinic acid; and polysaccharides such as dextrin.

[0029] The concentration of organic matter added to the medium as a carbon source should be adjusted appropriately for efficient growth. In general, the amount of addition can be selected from the range of 0.1 to 10 wt / vol %.

[0030] In addition to the above carbon sources, a nitrogen source can be added to the medium. As the nitrogen source, various nitrogen compounds that can be used in normal fermentation can be used. For example, inorganic nitrogen sources and organic nitrogen sources can be used. Examples of the inorganic nitrogen source include ammonium salts and nitrates. Preferred inorganic nitrogen sources are ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium hydrogen phosphate, potassium nitrate, and sodium nitrate. Examples of organic nitrogen sources include amino acids (glutamic acid, arginine, ornithine, etc.), fats and oils such as oleic acid, yeast extract, peptones (e.g., polypeptone N, soybean peptone, etc.), meat extracts (e.g., Ehrlich bonito extract, Lab-Remco powder, bouillon, etc.), seafood extracts, liver extracts, digested serum powders, fish oils, etc. More preferred are arginine, cysteine, citrulline, lysine, yeast extract, and peptones (e.g., polypeptone N, etc.).

[0031] Furthermore, in addition to carbon and nitrogen sources, the growth and activity may be enhanced by adding cofactors such as vitamins and inorganic compounds such as various salts to the medium. For example, the following are examples of microbial growth cofactors derived from plants and animals, such as inorganic compounds and vitamins:

[0032] Inorganic compounds Vitamins Potassium dihydrogen phosphate Biotin Magnesium Sulfate Folic Acid Manganese Sulfate Pyridoxine Sodium Chloride Thiamine Cobalt chloride Riboflavin Calcium chloride Nicotinic acid Zinc Sulfate Pantothenic Acid Copper Sulfate Vitamin B12 Alum Thioctic acid Sodium molybdate p-aminobenzoic acid Potassium chloride Vitamin K Boric acid etc. Nickel chloride Sodium Tungstate Sodium Selenite Ammonium Ferrous Sulfate Sodium Acetate Trihydrate Magnesium sulfate heptahydrate Manganese sulfate tetrahydrate

[0033] Methods for producing a medium by adding growth cofactors derived from animals and plants, such as inorganic compounds and vitamins, are known. The medium can be liquid, semi-solid, or solid. The preferred form of the medium is a liquid medium.

[0034] The medium of the present disclosure may also contain dextrins. By culturing anaerobic microorganisms in a medium containing dextrins, a solution containing a functional substance and dextrins can be obtained without adding dextrins even if dextrins are required in the culture solution after the culture. Dextrins can be added to the medium before and during the culture of the microorganisms. It can be done.

[0035] In addition, when culturing anaerobically, growth may be improved by adding reducing agents such as cysteine, cystine, sodium sulfide, sulfite, ascorbic acid, glutathione, thioglycolic acid, and rutin, or enzymes that break down reactive oxygen species, such as catalase and superoxide mutase, to the medium.

[0036] In the case of anaerobic cultivation, the gas phase and the aqueous phase during cultivation preferably do not contain air or oxygen, and may contain, for example, nitrogen and / or hydrogen in any ratio, or nitrogen and / or carbon dioxide in any ratio. These may be supplied in gaseous form. The proportion of hydrogen in the gas phase is not particularly limited, but is usually 0.5% or more and 100% or less, preferably 1.0% or more and 20% or less, and more preferably 2.0% or more and 10% or less.

[0037] The pH of the medium is preferably 5.0 or higher, more preferably 6.0 or higher, and even more preferably 6.5 or higher, while it is preferably 8.0 or lower, and more preferably 7.5 or lower. The culture temperature is preferably 20°C to 45°C, more preferably 25°C to 40°C, and further preferably 30°C to 37°C. The pressurization conditions for the culture vessel are not particularly limited as long as they allow growth, but examples of the pressurization conditions include a range of 0.001 to 1 MPa, and preferably 0.01 to 0.5 MPa. The culture time is usually 8 to 340 hours, preferably 12 to 170 hours, and more preferably 16 to 120 hours.

[0038] [(b1) process] The production method according to this embodiment includes a step (b1) of causing a microorganism capable of producing equol to produce equol from the isoflavone aglycone, and / or a step (b2) described below.

[0039] Examples of microorganisms capable of producing equol include microorganisms belonging to the genus Adlercreutzia, Atopobium, Bacteroides, Collinsella, Coriobacterium, Cryptobacterium, and Denitrobacterium. Living organisms, microorganisms belonging to the genus Eggerthella, microorganisms belonging to the genus Enterorhabdus, microorganisms belonging to the genus Eubacterium, and Microorganisms belonging to the genus Gordonibacter, microorganisms belonging to the genus Lactococcus, microorganisms belonging to the genus Olsenella, microorganisms belonging to the genus Paraeggerthella, microorganisms belonging to the genus Ruminococcus, microorganisms belonging to the genus Slackia Examples of microorganisms that can cause infection include those belonging to the genus Streptococcus.

[0040] Preferably, the microorganism is selected from the group consisting of Adlercreutzia equolifaciens subsp. equolifaciens, Adlercreutzia equolifaciens subsp. celatus, Bacteroides ovatus, and Eggerthella sp. Lactococcus galli, a microorganism belonging to the genus Eubacterium Microorganisms belonging to Lactococcus garvieae, Paraeggerthella sp., Ruminococcus productus Slackia isoflavoniconver Slackia equolifaciens, a microorganism belonging to the genus Slackia tens Microorganisms belonging to Slackia sp., Streptococcus It is a microorganism belonging to the genus Streptococcus intermedius.

[0041] More preferably, Adlercreutzia equolifaciens subsp. equolifaciens DSM 19450 strain. , Adlercreutzia equolifaciens subsp. celatus DSM 18785, Eggerthella sp. KCCM 10490, Lactococcus garvieae DSM 6783, Slackia isoflavoniconvertens DSM 22006, Examples thereof include Slackia equolifaciens DSM 24851 strain and Slackia sp. FERM AP-20729 strain. The above microorganisms may be used alone or in combination of two or more, regardless of genus, species or strain.

[0042] In addition, in this embodiment, the Adlercreutzia equolifaciens subsp. equolifaciens DSM 19450 strain is not limited to the same strain as the deposited strain, but may be a strain substantially equivalent to the deposited strain. The substantially equivalent strain refers to a strain that belongs to the same genus or species as the deposited strain and has the ability to produce equol. The substantially equivalent strain refers to a microorganism whose 16S rRNA gene base sequence has a homology of 97% or more, preferably 97.5% or more, more preferably 98% or more, even more preferably 98.7% or more, and even more preferably 99% or more with the 16S rRNA gene base sequence of the deposited strain. Furthermore, the microorganism capable of producing equol may be a strain bred from the deposited strain or a substantially equivalent strain by mutation treatment, genetic recombination, selection of natural mutants, or the like, so long as the effect of the present disclosure is not impaired. This similarly applies to the other deposited strains herein.

[0043] When the microorganism capable of producing equol produces β-glucosidase, the microorganism producing β-glucosidase may be the same as the microorganism capable of producing equol. In this case, steps (a) and (b1) are carried out successively in the same system.

[0044] The culture conditions may be those generally used in the culture of the above-mentioned microorganisms or appropriately modified culture conditions, and examples of the culture conditions include the same culture conditions as those for microorganisms that produce β-glucosidase.

[0045] [(b2) process] The production method according to this embodiment includes the step (b1) and / or the step (b2) of causing a microorganism capable of producing an equol derivative to produce an equol derivative from the isoflavone aglycone. An example of the equol derivative is 5-hydroxyequol.

[0046] When the equol derivative is 5-hydroxyequol, examples of microorganisms capable of producing 5-hydroxyequol include microorganisms belonging to the genera Adlercreutzia, Eggerthella, and Slackia. Microorganisms belonging to the genus Adlercreutzia include Adlercreutzia Microorganisms belonging to the genus Adlercreutzia equolifaciens are preferred, and However, Adlercreutzia equolifaciens subsp. equolifaciens DSM 19450 strain or Adlercreutzia Adlercreutzia equolifaciens subsp. celatus strain DSM 18785 is preferred.

[0047] Microorganisms belonging to the genus Eggerthella include microorganisms belonging to Eggerthella sp. Among them, Eggerthella sp. ) KCCM 10490 strain is preferred. Microorganisms belonging to the genus Slackia include Slackia isoflavonicum. Examples of microorganisms that can be used include those belonging to the genus Slackia isoflavoniconvertens, those belonging to Slackia equolifaciens, and those belonging to Slackia sp.

[0048] When the microorganism capable of producing an equol derivative produces β-glucosidase, the microorganism producing the β-glucosidase may be the same as the microorganism capable of producing an equol derivative. In this case, steps (a) and (b2) are carried out successively in the same system.

[0049] The culture conditions may be those generally used in the culture of the above-mentioned microorganisms or appropriately modified culture conditions, and examples of the culture conditions include the same culture conditions as those for microorganisms that produce β-glucosidase.

[0050] [(c) Process] The production method according to the present embodiment includes a step (c) of decomposing the glucose so that the content of the glucose in the composition is 0 g / L or more and less than 2.0 g / L, or 30 g / kg or less per solid content of the composition. Decomposing glucose refers to converting glucose into a compound having a smaller number of carbon atoms. The step (c) may be a step of decomposing the glucose by fermentation using a microorganism having sugar assimilation ability, or may be a step of decomposing the glucose using an enzyme that decomposes glucose.

[0051] When the step (c) utilizes fermentation using a microorganism capable of utilizing sugar, the microorganism capable of utilizing sugar is preferably one or more microorganisms selected from the group consisting of butyric acid bacteria, lactic acid bacteria, and yeast. More preferably, the microorganism is a microorganism belonging to the genus Clostridium. Microorganisms belonging to the genus Lactobacillus, microorganisms belonging to the genus Lactococcus, microorganisms belonging to the genus Leuconostoc, and microorganisms belonging to the genus Saccharomyces. One or more microorganisms selected from the group consisting of microorganisms belonging to the genus Saccharomyces be.

[0052] Microorganisms belonging to the genus Clostridium include Clostridium aeruginosa. Examples thereof include Clostridium asparagiforme strain DSM 15981 and Clostridium bolteae strain JCM 12243. As a microorganism belonging to the genus Lactobacillus, Lactobacillus brevis is Examples include Lactobacillus brevis DSM 20054 strain. Microorganisms belonging to the genus Lactococcus include Lactococcus lactis. Examples of such bacteria include Lactococcus lactis DSM 20481 strain. Microorganisms belonging to the genus Leuconostoc include Leuconostoc melanogaster. Examples include Leuconostoc mesenteroides subsp. mesenteroides DSM 20343 strain.

[0053] When a microorganism capable of sugar utilization produces β-glucosidase, the microorganism producing β-glucosidase may be the same as the microorganism capable of sugar utilization. In this case, steps (a) and (c) may be carried out successively in the same system.

[0054] The microorganism capable of sugar utilization may be of the same species as the microorganism capable of equol production. That is, the microorganism capable of sugar utilization may be a microorganism capable of equol production and sugar utilization. In this case, steps (b1) and (c) are carried out successively in the same system.

[0055] The microorganism capable of sugar utilization may be of the same species as the microorganism capable of producing an equol derivative. That is, the microorganism capable of sugar utilization may be a microorganism capable of producing an equol derivative and of sugar utilization. In this case, steps (b2) and (c) are carried out successively in the same system.

[0056] The culture conditions may be those generally used in the culture of the above-mentioned microorganisms or appropriately modified culture conditions, and examples of the culture conditions include the same culture conditions as those for microorganisms that produce β-glucosidase.

[0057] When the step (c) utilizes an enzyme that decomposes glucose, examples of the enzyme that decomposes glucose include glucose oxidase and glucose dehydrogenase. These may be used alone or in combination of two or more. Glucose oxidase and glucose dehydrogenase are enzymes that catalyze the reaction of oxidizing β-D-glucose to D-glucono-1,5-lactone.

[0058] The glucose oxidase and glucose dehydrogenase are not limited as long as they have the above enzyme activity, and may be derived from Aspergillus niger or Gluconobacter oxidase. Examples include those derived from Gluconobacter oxydans.

[0059] To decompose glucose using an enzyme that decomposes glucose, the enzyme may be added to a solution (medium) containing glucose. The amount of the enzyme added can be adjusted as appropriate, but is usually 1 U to 10,000 U, preferably 10 U to 5,000 U, more preferably 100 U to 2,000 U per 1 g of glucose.

[0060] The temperature at which glucose is decomposed is not particularly limited, but is usually 15°C or higher and 60°C or lower, preferably 25°C or higher and 50°C or lower, more preferably 30°C or higher and 40°C or lower, and particularly preferably 35°C. The pH at which glucose is decomposed is not particularly limited, but is usually 4 or more and 9 or less, preferably 4 or more and 7 or less, and more preferably 5 or more and 6 or less.

[0061] When steps (b1) and (b2) are performed, the order of these steps is not limited; step (b2) may be performed after step (b1), step (b2) may be performed after step (b1), or step (b1) and step (b2) may be performed simultaneously.

[0062] The order of step (b1) and / or step (b2) and step (c) is not limited; step (c) may be performed after step (b1) and / or step (b2), step (b1) and / or step (b2) may be performed after step (c), or step (b1) and / or step (b2) and step (c) may be performed simultaneously. Specific examples of the manner in which step (b1) and / or step (b2) are carried out simultaneously with step (c) include a manner in which an enzyme that decomposes glucose is added to a medium containing glucose, isoflavone aglycone, and a microorganism capable of producing equol, a manner in which a microorganism capable of producing equol and a microorganism capable of utilizing sugar are co-cultured in a medium containing glucose and isoflavone aglycone, and a manner in which a microorganism capable of producing equol and an microorganism capable of utilizing sugar are cultured in a medium containing glucose and isoflavone aglycone.

[0063] In step (c), the glucose content in the composition is 0 g / L or more and 2.0 g / L or less. The glucose is degraded to less than or equal to 30 g / kg of solids in the composition. In order to set the glucose content within the above range, the culture conditions and the conditions for the treatment with the glucose-degrading enzyme may be appropriately adjusted, for example by extending the culture time or the treatment time.

[0064] The glucose content in the composition is preferably 0 g / L or more and less than 2.0 g / L, more preferably 0 g / L or more and 1.0 g / L or less, even more preferably 0 g / L or more and 0.5 g / L or less, and particularly preferably 0 g / L or more and 0.2 g / L or less. In addition, the glucose content is preferably 30 g / kg or less per solid content of the composition, more preferably 10 g / kg or less, and even more preferably 4 g / kg or less. The solid content is determined by measuring the residual weight after drying the composition. Examples of drying methods include reduced pressure heating drying method, normal pressure heating drying method, and freeze drying method. The glucose content can be measured by an electrode method. When the composition does not contain water, such as when the composition is in a solid form, water can be added to the composition, glucose is dissolved in the water, and then the glucose content per solid content can be measured.

[0065] [Other steps] The manufacturing method of this embodiment may include, for example, a step of quantifying the amount of equol and / or equol derivative obtained. This can be done according to a conventional method. For example, a portion of the culture solution may be sampled, appropriately diluted, thoroughly stirred, and then filtered using a membrane such as a polytetrafluoroethylene (PTFE) membrane to remove insoluble matter, followed by quantifying the amount by high performance liquid chromatography (HPLC).

[0066] The production method of this embodiment may also include a step of recovering the obtained equol and / or equol derivative. The recovery step includes a purification step, a concentration step, and the like. Purification treatments in the purification step include sterilization of microorganisms by heat or the like; sterilization by microfiltration (MF), ultrafiltration (UF), and the like; removal of solids and polymeric substances; extraction with organic solvents, ionic liquids, and the like; and adsorption and decolorization using hydrophobic adsorbents, ion exchange resins, activated carbon columns, and the like. Concentration treatments in the concentration step include concentration using an evaporator, reverse osmosis membrane, and the like.

[0067] Furthermore, the obtained solution containing equol and / or an equol derivative can be powdered by freeze-drying, spray-drying, etc. In the powdering process, an excipient such as lactose, dextrin, corn starch, etc. can also be added.

[0068] [Method of producing food and drink] The composition produced by the production method according to the present embodiment can be suitably incorporated into food and drink, since it does not undergo the Maillard reaction and does not brown even when heated, either as is or in the presence of additional amino acids, etc. In this specification, food and drink includes supplements. That is, another embodiment of the present disclosure is a method for producing a food or beverage, comprising the following steps (a); (b1) and / or (b2); (c); and (d). (a) producing glucose and isoflavone aglycone from isoflavone glycoside; (b1) allowing a microorganism capable of producing equol to produce equol from the isoflavone aglycone; (b2) allowing a microorganism capable of producing an equol derivative to produce the equol derivative from the isoflavone aglycone; (c) decomposing the glucose so that the content of the glucose in a composition produced by a method including the steps (a), (b1) and / or (b2), and (c) is 0 g / L or more and less than 2.0 g / L, or 30 g / kg or less per solid content of the composition; and (d) a step of producing a food or beverage using a composition produced by a method including the steps (a), (b1) and / or (b2), and (c).

[0069] The food and drink produced by the production method of this embodiment contains equol and / or an equol derivative. The food and drink can be used as general food and drink, as well as specific health foods, nutritional supplements, functional foods, foods for the sick, food additives, etc. (including beverages). The form of the food and drink may be, for example, formed into a suitable form for consumption, such as granules, tablets, capsules, pastes, etc., using conventional means after adding appropriate auxiliary agents, and then provided for consumption. In addition, the composition may be added to various foods, such as processed meat foods such as ham and sausage, processed seafood foods such as kamaboko and chikuwa, bread, confectionery, butter, powdered milk, and fermented dairy products, or added to beverages such as water, fruit juice, milk, and soft drinks.

[0070] The above-mentioned food and drink may contain water, protein, carbohydrate, lipid, vitamins, minerals, organic acids, organic bases, fruit juice, flavors, etc. Examples of proteins include animal and vegetable proteins such as whole milk powder, skim milk powder, partially skim milk powder, casein, soy protein, egg protein, and meat protein, as well as hydrolysates thereof, and butter. Examples of carbohydrates include sugars, processed starch (dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), and dietary fiber. Examples of lipids include vegetable oils and fats such as lard, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, hydrogenated oils, and interesterified oils. Examples of vitamins include vitamin A, carotenes, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline, and folic acid. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium, and whey minerals. Examples of organic acids include malic acid, citric acid, lactic acid, and tartaric acid. Two or more of these components may be used in combination, and synthetic products and / or foods and beverages containing a large amount of these may be used.

[0071] The explanation of the method for producing a composition according to an embodiment of the present disclosure is incorporated herein by reference with respect to step (a), step (b1) and / or step (b2), and step (c). The method for producing a food or beverage according to the present embodiment may include the other steps described above.

[0072] (d) The step of producing a food or drink using the composition produced by the method including the steps (a), (b1) and / or (b2), and (c) can be carried out according to a conventional method. The amount, method and timing of incorporation of equol and / or equol derivatives can be appropriately selected. Furthermore, the food or drink can be enclosed in a container such as a bottle, bag, can, box or pack, as necessary.

[0073] [Fermentation composition] By using a method for producing a composition according to one embodiment of the present disclosure, it is possible to obtain a composition containing equol and / or an equol derivative in which the amount of glucose is reduced and which does not undergo the Maillard reaction even when heated, either alone or in the presence of additional amino acids, etc. That is, another embodiment of the present disclosure is a fermentation composition that contains equol and / or an equol derivative and is substantially free of glucose. Alternatively, another embodiment of the present disclosure is a fermentation composition that contains equol and / or an equol derivative and has a glucose content of 30 g / kg or less per solid content of the fermentation composition. An example of an equol derivative is 5-hydroxyequol. In addition, the fermented composition according to the present embodiment does not undergo the Maillard reaction and does not brown even when heated, either as is or in the presence of additional amino acids, etc., and therefore can be suitably included in foods and beverages. That is, the fermented composition can be used in foods and beverages.

[0074] "Substantially free of glucose" means that the glucose content is low enough that browning of the composition does not occur due to the Maillard reaction, and specifically includes cases where the glucose content is between 0 g / L and less than 2.0 g / L.

[0075] The glucose content in the fermentation composition is preferably 0 g / L or more and less than 2.0 g / L, more preferably 0 g / L or more and 1.0 g / L or less, even more preferably 0 g / L or more and 0.5 g / L or less, and particularly preferably 0 g / L or more and 0.2 g / L or less. The glucose content is preferably 30 g / kg or less per solid content of the fermentation composition, more preferably 10 g / kg or less, and even more preferably 4 g / kg or less. The solid content is determined by measuring the residual weight after drying the fermentation composition. Examples of drying methods include reduced pressure heating drying, normal pressure heating drying, and freeze drying. The glucose content can be measured by an electrode method. When the fermentation composition does not contain water, for example in a solid form, water can be added to the fermentation composition, glucose is dissolved in the water, and then the glucose content per solid content can be measured.

[0076] The content of equol and / or an equol derivative is preferably 1 g / kg or more, more preferably 5 g / kg or more, and even more preferably 10 g / kg or more, based on the solid content of the fermentation composition. The content of equol and / or equol derivatives can be measured by HPLC as described above.

[0077] The fermentation composition may include other components that may be included in the medium described above.

[0078] Foods and beverages containing the fermented composition can be used as general foods and beverages, as well as specific health foods, nutritional supplements, functional foods, foods for the sick, food additives, etc. (these also include beverages). As for the form of the food and beverage, for example, after adding an appropriate auxiliary agent, the food and beverage may be formed into an edible form, for example, granules, tablets, capsules, pastes, etc., using conventional means and provided for consumption, or may be added to various foods, for example, processed meat foods such as ham and sausage, processed seafood foods such as kamaboko and chikuwa, bread, confectionery, butter, powdered milk, fermented dairy products, or beverages such as water, fruit juice, milk, and soft drinks.

[0079] The above-mentioned food and drink may contain water, protein, carbohydrate, lipid, vitamins, minerals, organic acids, organic bases, fruit juice, flavors, etc. Examples of proteins include animal and vegetable proteins such as whole milk powder, skim milk powder, partially skim milk powder, casein, soy protein, egg protein, and meat protein, as well as hydrolysates thereof, and butter. Examples of carbohydrates include sugars, processed starch (dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), and dietary fiber. Examples of lipids include vegetable oils and fats such as lard, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, hydrogenated oils, and interesterified oils. Examples of vitamins include vitamin A, carotenes, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline, and folic acid. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium, and whey minerals. Examples of organic acids include malic acid, citric acid, lactic acid, and tartaric acid. Two or more of these components may be used in combination, and synthetic products and / or foods and beverages containing a large amount of these may be used.

[0080] The above-mentioned food and drink products can be produced according to conventional methods. The amount of the all derivative to be added, the method of addition, and the timing of addition can be appropriately selected. Furthermore, the above-mentioned food and drink can be enclosed in a container such as a bottle, a bag, a can, a box, or a pack, as necessary. EXAMPLES

[0081] The present disclosure will be described in more detail below with reference to specific examples, but the present disclosure is not limited to these examples.

[0082] (Method of measuring glucose concentration) 1 mL of the liquid to be measured was taken and the bacteria were precipitated by centrifugation. The supernatant was filtered through a 0.45 μm filter, and the filtrate was analyzed under the following conditions. If the concentration was outside the measurement range (10 mg / dL to 600 mg / dL), the filtrate was diluted with Milli-Q water and analyzed. Testing device: Glutest Neo Alpha manufactured by Sanwa Kagaku Kenkyusho Co., Ltd. Chip: Glutest Neo sensor manufactured by Sanwa Kagaku Kenkyusho Co., Ltd. The content per solid content (g / kg) was calculated by measuring the solid content (kg / L) of the liquid to be measured using an electronic moisture meter (Shimadzu MOC-120H) and dividing the glucose concentration (g / L) by the solid content.

[0083] (Method of measuring equol concentration and equol derivatives) 20 μL of the liquid to be measured was taken and diluted 50 times with a diluent consisting of ethanol: Milli-Q water = 70:30 (v / v). The diluted liquid was filtered through a 0.45 μm filter, and the filtrate was analyzed under the following HPLC conditions. The equol standard used was (S)-equol manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., and the 5-hydroxyequol standard used was 5-hydroxyequol manufactured by Toronto Research Chemicals. {HPLC conditions} Column: Phenomenex SYNERGI, 4μm, POLAR-R, 150mm×4.6mm Eluent: distilled water / methanol = 55 / 45 (v / v) Temperature: 40℃ Detection wavelength: 280nm Flow rate: 1.0mL / min Injection: 10μL Time: 30 minutes

[0084] [Test Example 1] (Preparation of pre-culture medium) Dissolve 35.4g of Thermo Scientific Anaerobe Basal Broth (ABB) in 1L of water. The pre-culture medium was prepared by dispensing 10 mL of the pre-culture medium into a test tube, which was then sealed with a butyl rubber stopper. After replacing the gas with nitrogen gas, the tube was sterilized in an autoclave.

[0085] (Enzyme treatment) 0.16 g / L of Pectinase G Amano (Amano Enzyme Co., Ltd.) was added to 16 g / L of soybean hypocotyl extract (containing 80% isoflavone glycosides such as daidzin, glycitin, and genistin), and the mixture was kept at 50°C overnight with stirring to release sugars from the isoflavone glycosides.

[0086] (Preparation of main culture medium) The final concentration of the enzyme treatment solution was 35.4 g / L of ABB medium, 1 g / L of arginine, and β-cysteine. Chlorodextrin was added to the solution so that the concentration was 16 g / L, and the solution was dispensed in 1 L portions into 2 L mini jars. After replacing the atmosphere with nitrogen gas, the solution was sterilized in an autoclave.

[0087] (preculture) The preculture medium was inoculated with the microorganisms in the combinations shown in Table 1, and then the atmosphere was replaced with anaerobic gas and cultured at 37°C and 200 spm for 2 days. Adlercreutzia equolifaciens subsp. celatus (DSM 18785) is a microorganism capable of producing equol. The strains used were either the Adlercreutzia equolifaciens DSM 19450 strain (referred to as AC in the table) or the Adlercreutzia equolifaciens DSM 19450 strain (referred to as AE in the table), both of which are capable of producing equol derivatives (especially 5-hydroxyequol). In addition, the following microorganisms capable of sugar utilization were used: Clostridium asparagiforme DSM 15981 strain (represented as CA in the table), Clostridium bolteae JCM 12243 strain (represented as CB in the table), Lactobacillus brevis DSM 20054 strain (represented as LB in the table), Lactococcus lactis DSM 20481 strain (represented as LL in the table), and Leuconostoc mesenteroides subsp. mesenteroides DSM 20343 strain (represented as LM in the table).

[0088] (main culture) After inoculating the main culture medium with the above-mentioned preculture medium, the main culture medium was cultured at 37°C and 500 rpm for 2 days while aerating with anaerobic gas through a 0.22 μm filter.

[0089] (result) After cultivation, the equol concentration, 5-hydroxyequol concentration, and glucose concentration were measured. The results are shown in Table 1. The glucose concentration expressed in "g / kg" indicates the glucose content per solid content.

[0090] [Table 1]

[0091] It can be seen from Comparative Examples 1 and 2 and Examples 1 to 6 that equol and 5-hydroxyequol were produced by culturing microorganisms capable of producing equol. Furthermore, it can be seen from Examples 1 to 6 that sugar was assimilated by co-cultivating microorganisms capable of assimilating sugar, with no glucose remaining in the culture medium.

[0092] [Test Example 2] (Preparation of pre-culture medium) A preculture medium was prepared in the same manner as in Test Example 1.

[0093] (Enzyme treatment) 0.16 g / L of Pectinase G Amano was added to 16 g / L of soybean hypocotyl extract (containing 80% isoflavone glycosides such as daidzin, glycitin, and genistin), and the mixture was kept at 50°C overnight with stirring to release sugars from the isoflavone glycosides.

[0094] (Preparation of main culture medium) The final concentration of the enzyme treatment solution was 35.4 g / L of ABB medium, 1 g / L of arginine, and β-cysteine. Chlorodextrin was added to the solution so that the concentration was 16 g / L, and 50 mL of the solution was dispensed into vials, which were then sealed with butyl rubber stoppers. After replacing the atmosphere with nitrogen gas, the solution was sterilized in an autoclave.

[0095] (preculture) The microorganisms capable of producing equol shown in Table 2 were inoculated into the pre-culture medium, and the atmosphere was replaced with anaerobic gas, followed by cultivation at 37°C and 200 spm for 2 days.

[0096] (main culture) After inoculating the main culture medium with the above preculture, the atmosphere was replaced with anaerobic gas and cultured at 37°C and 200 spm for 3 days. The specimens cultured as described above were designated Comparative Examples 3 and 4, and the equol concentration, 5-hydroxyequol concentration, and glucose concentration were measured.

[0097] (Yeast treatment) To Comparative Examples 3 and 4, 1 g of baker's yeast (Super Camellia Dry Yeast manufactured by Nisshin Seifun Welna Co., Ltd.) was added per 100 mL of culture solution, and the mixture was stored overnight at 30° C. to prepare Examples 7 and 8, respectively.

[0098] (result) The measurement results of equol concentration, 5-hydroxyequol concentration, and glucose concentration in Comparative Examples 3 and 4 and Examples 7 and 8 are shown in Table 2.

[0099] [Table 2]

[0100] The glucose concentration in Examples 7 and 8 after yeast treatment was significantly lower than that in Comparative Examples 3 and 4 after main culture. It is believed that glucose was assimilated by the yeast. In addition, the yeast-treated samples had a characteristic yeast smell and a good flavor.

[0101] [Test Example 3] The solutions of Comparative Example 3 and Example 7 were centrifuged to remove the bacterial cells and insoluble components, and the supernatants were collected. The absorbance of the supernatants was measured at 420 nm. In addition, after heating at 80° C. for 2 hours, the absorbance was measured in the same manner. The results are shown in Table 3.

[0102] [Table 3]

[0103] In Comparative Example 3, the absorbance increased due to heating. This is believed to be because the remaining glucose caused the Maillard reaction, resulting in browning. On the other hand, in Example 7, the glucose concentration was reduced to 0.2 g / L or less due to the action of yeast, so no increase in absorbance was observed even after heat treatment.

[0104] [Test Example 4] Glycine was added to the solutions of Comparative Example 3 and Example 7 to a concentration of 2 g / L, and then the solution was centrifuged to remove the cells and insoluble components, and the supernatant was collected. The absorbance of the supernatant was measured at 420 nm. Similarly, glycine was added to the solutions of Comparative Example 3 and Example 7 to a concentration of 2 g / L, and the solution was heated at 80° C. for 2 hours, and the absorbance was measured in the same manner. The results are shown in Table 4.

[0105] [Table 4]

[0106] In Comparative Example 3, the absorbance increased due to heating. This is thought to be because the Maillard reaction proceeded due to the remaining glucose and the added glycine, causing browning. On the other hand, in Example 7, the glucose concentration was reduced to 0.2 g / L or less due to the action of yeast, so no increase in absorbance was confirmed even after the addition of glycine after the heat treatment.

[0107] [Test Example 5] Main culture was carried out in the same manner as in Test Example 2. After culture, the production of equol and 5-hydroxyequol was confirmed by HPLC analysis, and the glucose concentration was measured. After confirming that glucose remained, commercially available glucose oxidase (Amano Enzyme Inc.) and peroxidase (Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stored overnight at 30°C. The equol concentration, 5-hydroxyequol concentration, and glucose concentration were then measured. The culture solutions before the enzyme addition treatment are shown in Table 5 as Comparative Examples 5 and 6, and the culture solutions after the enzyme addition treatment are shown as Examples 9 and 10.

[0108] It can be seen that the glucose is decomposed by the enzyme addition treatment and does not remain in the solution after treatment.

[0109] [Table 5]

[0110] [Test Example 6] (Preparation of pre-culture medium) A preculture medium was prepared in the same manner as in Test Example 1. (Enzyme treatment) 0.16 g / L of Pectinase G Amano (Amano Enzyme Co., Ltd.) was added to 16 g / L of soybean hypocotyl extract (containing 44% isoflavone glycosides such as daidzin, glycitin, and genistin), and the mixture was kept at 50°C overnight with stirring to release sugars from the isoflavone glycosides.

[0111] (Preparation of main culture medium) The final concentration of the enzyme treatment solution was 35.4 g / L of ABB medium, 1 g / L of arginine, and β-cysteine. Chlorodextrin was added to the solution so that the concentration was 16 g / L, and the solution was dispensed in 1 L portions into 2 L mini jars. After replacing the atmosphere with nitrogen gas, the solution was sterilized in an autoclave.

[0112] (preculture) The microorganisms were inoculated into the preculture medium in the combination shown in Table 5, and then the atmosphere was replaced with anaerobic gas and cultured at 37° C. and 200 spm for 2 days. The microorganisms used were the same as those in Test Example 1. (main culture) After inoculating the main culture medium with the above-mentioned preculture medium, the main culture medium was cultured at 37°C and 500 rpm for 2 days while aerating with anaerobic gas through a 0.22 μm filter. (result) After incubation, the equol concentration, 5-hydroxyequol concentration, and glucose concentration were measured. The results are shown in Table 6.

[0113] [Table 6]

[0114] It can be seen from Comparative Examples 7 and 8 and Examples 11 to 16 that equol and 5-hydroxyequol were produced by culturing microorganisms capable of producing equol. Furthermore, it can be seen from Examples 11 to 16 that sugar was assimilated by co-cultivating microorganisms capable of assimilating sugar, with no glucose remaining in the culture medium.

[0115] [Test Example 7] Commercially available glucose oxidase (Amano Enzyme Inc.) and peroxidase (Fujifilm Wako Pure Chemical Industries, Ltd.) were added to Comparative Examples 7 and 8, and the mixture was stored overnight at 30° C. Thereafter, the equol concentration, 5-hydroxyequol concentration, and glucose concentration were measured.

[0116] (result) The culture solutions after the enzyme addition treatment are designated as Examples 17 and 18, and the measurement results of the equol concentration, 5-hydroxyequol concentration, and glucose concentration in Comparative Examples 7 and 8 and Examples 17 and 18 are shown in Table 7.

[0117] [Table 7]

[0118] It can be seen that the glucose is decomposed by the enzyme addition treatment and does not remain in the solution after treatment.

[0119] [Test Example 8] The solutions of Comparative Example 7 and Example 17 were centrifuged to remove the bacterial cells and insoluble components, and the supernatants were collected. The absorbance of the supernatants was measured at 420 nm. In addition, after heating at 80° C. for 2 hours, the absorbance was measured in the same manner. The results are shown in Table 8.

[0120] [Table 8]

[0121] In Comparative Example 7, the absorbance increased due to heating. This is believed to be because the remaining glucose caused the Maillard reaction, resulting in browning. On the other hand, in Example 17, the glucose concentration was reduced to 0.1 g / L or less due to the action of the enzyme, so no increase in absorbance was observed even after heat treatment.

Claims

[Claim 1] A method for producing a composition, comprising the following steps (a); (b1) and / or (b2); and (c). (a) producing glucose and isoflavone aglycone from isoflavone glycoside; (b1) allowing a microorganism capable of producing equol to produce equol from the isoflavone aglycone; (b2) allowing a microorganism capable of producing an equol derivative to produce the equol derivative from the isoflavone aglycone; and (c) decomposing the glucose so that the content of the glucose in the composition is 0 g / L or more and less than 2.0 g / L, or 30 g / kg or less per solid content of the composition;

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