Method for producing dealkylated polyphenol

Abstract

To provide at least a technique for producing dealkylated polyphenol.SOLUTION: Provided is a method for producing a dealkylated polyphenol, comprising the step (a1). In step (a1), in a solution containing a polyphenol having an alkoxy group in its side chain, a dealkylated polyphenol in which the alkyl group of the alkoxy group is eliminated from the polyphenol having an alkoxy group in the side chain is produced by a microorganism belonging to the genus Acetobacterium and having the ability to eliminate the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain to produce the dealkylated polyphenol.SELECTED DRAWING: None

Description

Technical Field

[0001] The present disclosure relates to a method for producing dealkylated polyphenols. Specifically, it relates to a method for producing dealkylated polyphenols in which the alkyl group of an alkoxy group has been eliminated from a polyphenol having an alkoxy group in a side chain.

Background Art

[0002] A variety of polyphenols are present in plants. It has become clear that when humans ingest these, various functions are exhibited, such as antioxidant, antiglycation, anti-inflammatory, antidiabetic, antihyperlipidemic, hormone replacement, and improvement of brain functions such as anti-amnesia. Polyphenols are also said to be the seventh nutrient after the five major nutrients and dietary fiber (Non-Patent Document 1). Polyphenols include lignans such as curcumin, sesamin, and SECO; phenolic acids such as ferulic acid; xanthones such as capsaicin and mangostin; and flavonoids such as anthocyanidins (such as petunidin), flavanones (such as hesperetin, nobilitin, and isoxanthohumol), chalcones (such as xanthohumol), and isoflavones (such as glycitein and biochanin). Among polyphenols, there are those having an alkoxy group such as a methoxy group or an ethoxy group. By the dealkoxylation of these, an improvement in solubility in water, an improvement in absorption rate into the living body, an increase in functionality, etc. are expected.

[0003] For example, 8-prenylnaringenin obtained by demethylation of isoxanthohumol (a type of flavanone) is known to have estrogen-like activity and activity for suppressing disuse muscle atrophy (Patent Document 1). In addition, 6-hydroxy daidzein, which is a demethylated product of glycitein (a type of isoflavone), can serve as a raw material for equol, which has estrogen-like activity (Patent Document 2). Since equol has strong physiological effects similar to female hormones, its use for preventing and improving menopausal symptoms and osteoporosis (Patent Document 3), preventing and treating skin aging and wrinkles (Patent Document 4), alleviating allergic symptoms (Patent Document 5), etc. has been proposed. Against these backgrounds, there is a demand for the development of an industrial production method that uses polyphenols containing an alkoxy group in the side chain in plants as raw materials, and eliminates the alkyl group of the alkoxy group from the polyphenols to produce dealkylated polyphenols.

[0004] As such a production method, a production method using microorganisms has been developed. For example, it has been reported that Blautia sp. MRG-PMF1 strain demethylates polymethoxyflavo nes (Non-Patent Document 2). In addition, it has been reported that Eubacterium limosum ATCC 8486 strain and Blautia producta ATCC 27340 strain (former Peptostreptococcus productus ATCC 27340 strain) demethylate isoxanthohumol to produce 8-prenylnaringenin (Patent Document 6, Patent Document 7). In addition, it has been reported that Blautia coccoides JCM 1395 strain, Blautia schinkii DSM 10518 strain, and microorganisms belonging to Eubacterium limosum demethylate glycitein to produce 6-hydroxy daidzein (Patent Document 2, Patent Document 8).

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2013-35811 [Patent Document 2] Japanese Patent Application Laid-Open No. 2020-058319 [Patent Document 3] Japanese Patent Publication No. 2001-523258 [Patent Document 4] Japanese Patent Publication No. 2002-511860 [Patent Document 5] Specification of Patent No. 4479505 [Patent Document 6] Japanese Patent Publication No. 2008-532558 [Patent Document 7] Japanese Patent Application Laid-Open No. 2020-115858 [Patent Document 8] Japanese Patent Application Laid-Open No. 2010-104241 [Non-Patent Document]

[0006] [Non-Patent Document 1] Chemistry and Biology, 53, 442-448 (2015) [Non-Patent Document 2] S. Burapan, et al., Journal of Agric. Food Chem., 65, 1620-1629 (2017) [Summary of the Invention] [Problems to be Solved by the Invention]

[0007] The problem of the present disclosure is to provide at least a technique for producing dealkylated polyphenols. Preferably, it is to provide a technique for producing dealkylated flavonoids. [Means for Solving the Problems]

[0008] <1>A method for producing a dealkylated flavonoid, comprising the following step (a1). Step (a1): In a solution containing a flavonoid having an alkoxy group in the side chain, a microorganism belonging to the genus Acetobacterium having the ability to eliminate the alkyl group of the alkoxy group of the flavonoid having an alkoxy group in the side chain to produce a dealkylated flavonoid is used to produce a dealkylated flavonoid in which the alkyl group of the alkoxy group has been eliminated from the flavonoid having an alkoxy group in the side chain. <2>The production method according to <1>, wherein the alkoxy group is a methoxy group and the alkyl group is a methyl group. <3>The production method according to <1> or <2>, wherein the flavonoid is flavanone and the dealkylated flavonoid is dealkylated flavanone. <4>The production method according to <1> or <2>, wherein the flavonoid is isoflavone and the dealkylated flavonoid is dealkylated isoflavone. <5>The production method according to <3>, wherein the flavanone is isoxanthohumol, the dealkylated flavanone is 8-prenylnaringenin, the alkoxy group is a methoxy group, and the alkyl group is a methyl group. <6>The production method according to <4>, wherein the isoflavone is glycitein, the dealkylated isoflavone is 6-hydroxy daidzein, the alkoxy group is a methoxy group, and the alkyl group is a methyl group. <7>The production method according to any one of <1> to <6>, wherein the microorganism belonging to the genus Acetobacterium is a microorganism belonging to Acetobacterium bakii. <8>The production method according to <7>, wherein the microorganism belonging to Acetobacterium bakii is Acetobacterium bakii DSM 8239 strain.

Advantages of the Invention

[0009] The present disclosure can at least achieve the effect of providing a technique for producing dealkylated polyphenols, and can also achieve the effect of providing useful microorganisms therefor. Preferably, it can achieve the effect of providing a technique for producing dealkylated flavonoids. This technique utilizes a conversion reaction (fermentation) by microorganisms. By using this technique, dealkylated polyphenols can be efficiently produced from polyphenols having an alkoxy group in the side chain .

Mode for Carrying Out the Invention

[0010] Each configuration and their combinations in each embodiment are examples, and within the scope not departing from the gist of the present disclosure, additions, omissions, substitutions, and other changes to the configuration can be made as appropriate. The present disclosure is not limited by the embodiments, but is limited only by the scope of the claims

[0011] In the present disclosure, microorganisms assigned with JCM numbers are microorganisms stored in Japan Collection of Microorganisms (Microbial Material Development Office, National Institute of Advanced Industrial Science and Technology, Postal Code : 305 - 0074, Address: 3 - 1 - 1, Takano - dai, Tsukuba City, Ibaraki Prefecture), and are microorganisms that can be obtained from the same institution . Also, microorganisms assigned with DSM numbers are microorganisms stored in DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Address: Inhoffenstraβe 7B, 38124 Braunschweig, Germany), and are microorganisms that can be obtained from the same institution

[0012] <Method for Producing Dealkylated Polyphenol> One aspect of the present disclosure is a method for producing dealkylated polyphenols, including the following step (a1). Step (a1): In a solution containing a polyphenol having an alkoxy group in a side chain, a microorganism belonging to the genus Acetobacterium having the ability to eliminate the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain to generate a dealkylated polyphenol is used to generate a dealkylated polyphenol in which the alkyl group of the alkoxy group has been eliminated from the polyphenol having an alkoxy group in the side chain.

[0013] (Polyphenol having an alkoxy group in a side chain) The polyphenol having an alkoxy group in the side chain may have one alkoxy group or may have a plurality of alkoxy groups. In any case, its raw material is not particularly limited.

[0014] Examples of the alkoxy group in the polyphenol having an alkoxy group in the side chain include a methoxy group, an ethoxy group, a propoxy group, a butoxy group, etc. Preferably, it is a methoxy group.

[0015] Examples of the polyphenol having an alkoxy group in the side chain include phenolic acids having an alkoxy group in the side chain, lignans having an alkoxy group in the side chain, chromans having an alkoxy group in the side chain, coumarins having an alkoxy group in the side chain, flavonoids having an alkoxy group in the side chain, xanthones having an alkoxy group in the side chain, simple phenols having an alkoxy group in the side chain, etc.

[0016] Examples of the phenolic acid having an alkoxy group in the side chain include ferulic acid (the alkoxy group is one methoxy group), anisic acid (the alkoxy group is one methoxy group), vanillic acid (the alkoxy group is one methoxy group), syringic acid (the alkoxy group is two methoxy groups), etc.

[0017] Examples of the lignan having an alkoxy group in the side chain include pinoresinol (the alkoxy group is two methoxy groups), secoisolariciresinol (the alkoxy group is two methoxy groups), and the like.

[0018] Examples of the chroman having an alkoxy group in the side chain include 6-methoxychroman ( the alkoxy group is one methoxy group), 2-methoxychroman (the alkoxy group is one methoxy group), 5-methoxychroman (the alkoxy group is one methoxy group), and the like.

[0019] Examples of the coumarin having an alkoxy group in the side chain include scopoletin (the alkoxy group is two methoxy groups), scopolin (the alkoxy group is one methoxy group), isoscopolin (the alkoxy group is one methoxy group), and the like.

[0020] Examples of the flavonoid having an alkoxy group in the side chain include anthocyanidin having an alkoxy group in the side chain, flavan having an alkoxy group in the side chain, flavanol having an alkoxy group in the side chain (sometimes referred to as "catechin having an alkoxy group in the side chain"), flavone having an alkoxy group in the side chain, flavonol having an alkoxy group in the side chain, flavanone having an alkoxy group in the side chain, isoflavone having an alkoxy group in the side chain, chalcone having an alkoxy group in the side chain, and the like.

[0021] Examples of the anthocyanidin having an alkoxy group in the side chain include malvidin (the alkoxy group is two methoxy groups), peonidin (the alkoxy group is one methoxy group), and the like.

[0022] Examples of flavans having an alkoxy group in the side chain include 4'-methoxyflavan (the alkoxy group is one methoxy group), 3'-methoxyflavan (the alkoxy group is one methoxy group), 7-methoxyflavan (the alkoxy group is one methoxy group), and the like.

[0023] Examples of flavanols having an alkoxy group in the side chain include 3'-O-methylcatechin (the alkoxy group is one methoxy group), 4'-O-methylepicatechin (the alkoxy group is one methoxy group), 4'-O-methylepigallocatechin (the alkoxy group is one methoxy group), and the like.

[0024] Examples of flavones having an alkoxy group in the side chain include nobilitin (the alkoxy group is six methoxy groups), sinensetin (the alkoxy group is five methoxy groups), tangeretin (the alkoxy group is five methoxy groups), ougonin (the alkoxy group is one methoxy group), and the like.

[0025] Examples of flavonols having an alkoxy group in the side chain include patuletin (the alkoxy group is one methoxy group), tamarixetin (the alkoxy group is one methoxy group), syringetin (the alkoxy group is two methoxy groups), izalpinin (the alkoxy group is one methoxy group), and the like.

[0026] Examples of flavanones having an alkoxy group in the side chain include isoxanthohumol (the alkoxy group is one methoxy group), hesperetin (the alkoxy group is one methoxy group), and the like.

[0027] Examples of the isoflavone having an alkoxy group in the side chain include glycitein (the alkoxy group is one methoxy group), biochanin A (the alkoxy group is one methoxy group), formononetin (the alkoxy group is one methoxy group), tectigenin (the alkoxy group is one methoxy group), and the like.

[0028] Examples of the chalcone having an alkoxy group in the side chain include xanthohumol (the alkoxy group is one methoxy group), and the like.

[0029] Examples of the xanthone having an alkoxy group in the side chain include α-mangostin (the alkoxy group is one methoxy group), β-mangostin (the alkoxy group is two methoxy groups), and the like.

[0030] Examples of the simple phenols having an alkoxy group in the side chain include paeonol (the alkoxy group is one methoxy group), anisole (the alkoxy group is one methoxy group), and the like.

[0031] (Dealkylated polyphenol) In the present disclosure, the polyphenol generated by the elimination of the alkyl group of the alkoxy group from the "polyphenol having an alkoxy group in the side chain" by the step (a1) may be referred to as a "dealkylated polyphenol". At this time, the alkoxy group and the alkyl group eliminated from the alkoxy group are such that when the alkoxy group is a methoxy group, the alkyl group is a methyl group; when the alkoxy group is an ethoxy group, the alkyl group is an ethyl group; when the alkoxy group is a propoxy group, the alkyl group is a propyl group; and when the alkoxy group is a butoxy group, the alkyl group is a butyl group. Preferably, the alkoxy group is a methoxy group and the alkyl group is a methyl group.

[0032] In the present disclosure, when a dealkylated polyphenol is generated from a polyphenol having one alkoxy group in a side chain, it is generated by the elimination of the alkyl group of the one alkoxy group. When it is generated from a polyphenol having a plurality of alkoxy groups in a side chain, it may be generated by the elimination of the alkyl group of one alkoxy group among the plurality of alkoxy groups, or it may be generated by the elimination of the alkyl groups of some of the plurality of alkoxy groups (not one alkoxy group and not all alkoxy groups), or it may be generated by the elimination of the alkyl groups of all of the plurality of alkoxy groups. In addition, a polyphenol having a plurality of alkoxy groups in a side chain may have all of the plurality of alkoxy groups as the same alkoxy group or different alkoxy groups. In addition, when the alkyl groups of a plurality of alkoxy groups (not one alkoxy group and not all alkoxy groups) are eliminated from a polyphenol having a plurality of alkoxy groups, alkoxy groups remain in the generated dealkylated polyphenol. Therefore, the generated dealkylated polyphenol can be used as the "polyphenol having an alkoxy group in a side chain". For example, when the alkyl group of one alkoxy group is eliminated from a polyphenol having three alkoxy groups, two alkoxy groups remain in the generated dealkylated polyphenol. Therefore, the generated dealkylated polyphenol can be used as the "polyphenol having an alkoxy group in a side chain".

[0033] In addition, the fact that a polyphenol generated by the elimination of the alkyl group of the alkoxy group from the "polyphenol having an alkoxy group in a side chain" described above may be referred to as a "dealkylated polyphenol" also applies to specific examples of the "polyphenol having an alkoxy group in a side chain". For example, a product generated by the elimination of the alkyl group of the alkoxy group from a "phenolic acid having an alkoxy group in a side chain" may be referred to as a "dealkylated phenolic acid". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "lignan having an alkoxy group in the side chain" may be referred to as "dealkylated lignan". Similarly, from "chroman having an alkoxy group in the side chain", the alkyl group of the alkoxy group is eliminated to form a product that may be referred to as "dealkylated chroman". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "coumarin having an alkoxy group in the side chain" may be referred to as "dealkylated coumarin". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "flavonoid having an alkoxy group in the side chain" may be referred to as "dealkylated flavonoid". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "anthocyanidin having an alkoxy group in the side chain" may be referred to as "dealkylated anthocyanidin". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "flavan having an alkoxy group in the side chain" may be referred to as "dealkylated flavan". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "flavanol having an alkoxy group in the side chain" may be referred to as "dealkylated flavanol". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "flavone having an alkoxy group in the side chain" may be referred to as "dealkylated flavone". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "flavonol having an alkoxy group in the side chain" may be referred to as "dealkylated flavonol". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "flavanone having an alkoxy group in the side chain" may be referred to as "dealkylated flavanone". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "isoflavone having an alkoxy group in the side chain" may be referred to as "dealkylated isoflavone". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "chalcone having an alkoxy group in the side chain" may be referred to as "dealkylated chalcone". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "xanthone having an alkoxy group in the side chain" may be referred to as "dealkylated xanthone". Similarly, a product formed by the elimination of the alkyl group of the alkoxy group from "simple phenols having an alkoxy group in the side chain" may be referred to as "dealkylated simple phenols".

[0034] Examples of the dealkylated phenolic acid include, for example, caffeic acid formed by the elimination of the methyl group of one methoxy group of ferulic acid (the alkoxy group is one methoxy group), salicylic acid formed by the elimination of the methyl group of one methoxy group of anisic acid (the alkoxy group is one methoxy group), protocatechuic acid formed by the elimination of the methyl group of one methoxy group of vanillic acid (the alkoxy group is one methoxy group), 3-O-methylgallate formed by the elimination of the methyl group of one methoxy group of syringic acid (the alkoxy group is two methoxy groups), gallic acid formed by the elimination of the methyl groups of two methoxy groups, and the like.

[0035] Examples of the dealkylated lignan include, for example, 1,2-benzenediol formed by the elimination of the methyl group of one methoxy group of pinoresinol (the alkoxy group is two methoxy groups), 3,3'-bisdemethylpinoresinol formed by the elimination of the methyl groups of two methoxy groups, O-demethylsecoisolariciresinol formed by the elimination of the methyl group of one methoxy group of secoisolariciresinol (the alkoxy group is two methoxy groups), dihydroxyenterodiol formed by the elimination of the methyl groups of two methoxy groups, and the like.

[0036] Examples of the dealkylated chromane include, for example, 6-hydroxy chromane formed by the elimination of the methyl group of one methoxy group of 6-methoxy chromane (wherein the alkoxy group is one methoxy group). 2-hydroxy chromane formed by the elimination of the methyl group of one methoxy group of 2-methoxy chromane (wherein the alkoxy group is one methoxy group). 5-hydroxy chromane formed by the elimination of the methyl group of one methoxy group of 5-methoxy chromane (wherein the alkoxy group is one methoxy group), and the like.

[0037] Examples of the dealkylated coumarin include, for example, Scopletin, isoscopletin formed by the elimination of the methyl group of one methoxy group of scopoletin (wherein the alkoxy group is two methoxy groups), and esculetin formed by the elimination of the methyl groups of two methoxy groups. Esculetin formed by the elimination of the methyl group of one methoxy group of scopletin (wherein the alkoxy group is one methoxy group). Esculetin formed by the elimination of the methyl group of one methoxy group of isoscopletin (wherein the alkoxy group is one methoxy group), and the like.

[0038] Examples of the dealkylated anthocyanidin include, for example, Petunidin formed by the elimination of the methyl group of one methoxy group of malvidin (wherein the alkoxy group is two methoxy groups), and delphinidin formed by the elimination of the methyl groups of two methoxy groups. Delphinidin formed by the elimination of the methyl group of one methoxy group of peonidin (wherein the alkoxy group is one methoxy group), and the like.

[0039] Examples of the dealkylated flavan include, for example, 4'-Hydroxyflavan formed by the elimination of the methyl group of one methoxy group of 4'-methoxyflavan (wherein the alkoxy group is one methoxy group). 3'-Hydroxyflavan formed by the elimination of the methyl group of one methoxy group of 3'-methoxyflavan (wherein the alkoxy group is one methoxy group). Examples include 7-hydroxyflavan formed by the elimination of the methyl group of one methoxy group of 7-methoxyflavan (wherein the alkoxy group is one methoxy group).

[0040] Examples of the dealkylated flavanol include, for example, Catechin formed by the elimination of the methyl group of one methoxy group of 3'-O-methylcatechin (wherein the alkoxy group is one methoxy group). Epicatechin formed by the elimination of the methyl group of one methoxy group of 4'-O-methylepicatechin (wherein the alkoxy group is one methoxy group). Examples include epigallocatechin formed by the elimination of the methyl group of one methoxy group of 4'-O-methyl epigallocatechin (wherein the alkoxy group is one methoxy group).

[0041] Examples of the dealkylated flavone include, for example, Among nobilitin (wherein the alkoxy group is six methoxy groups), 4'-demethylnobilitin formed by the elimination of the methyl group of one methoxy group, 3',4'-dimethylnobilitin formed by the elimination of the methyl groups of two methoxy groups, sideritoflavone formed by the elimination of the methyl groups of three methoxy groups, leucopelargonidin formed by the elimination of the methyl groups of four methoxy groups, 2-(3,4-dihydroxyphenyl)-5,6,8-trihydroxy-7-methoxy-4H-1-benzopyran-4-one formed by the elimination of the methyl groups of five methoxy groups, and vitagenin formed by the elimination of the methyl groups of six methoxy groups. Among sinensetin (wherein the alkoxy group is five methoxy groups), 4'-desmethylsinensetin formed by the elimination of the methyl group of one methoxy group, 3',4'-dihydroxy-5,6,7-trimethoxyflavone formed by the elimination of the methyl groups of two methoxy groups, 3',4',7'-trihydro xy-5,6-dimethylxyflavone formed by the elimination of the methyl groups of three methoxy groups, karajuflavone formed by the elimination of the methyl groups of four methoxy groups, 6-hydroxyltheolin formed by the elimination of the methyl groups of five methoxy groups, Among tangeretin (wherein the alkoxy group is five methoxy groups), 4'-hydroxy-5,6,7,8-tetramethoxyflavone formed by the elimination of the methyl group of one methoxy group, xanthomicrol formed by the elimination of the methyl groups of two methoxy groups, isothymusin formed by the elimination of the methyl groups of three methoxy groups, 5,7,8-trihydroxy-2-(4-hydroxyphenyl)-6-methoxy-4H-1-benzopyran-4-one formed by the elimination of the methyl groups of four methoxy groups, nor-tangeretin formed by the elimination of the methyl groups of five methoxy groups, Examples include nor-ogonin formed by the elimination of the methyl group of one methoxy group of oogonin (wherein the alkoxy group is one methoxy group).

[0042] Examples of the dealkylated flavonol include, for example, Kelsegetin formed by the elimination of the methyl group of one methoxy group of patuletin (wherein the alkoxy group is one methoxy group). Quercetin formed by the elimination of the methyl group of one methoxy group of tamarixetin (wherein the alkoxy group is one methoxy group). Among syringetin (wherein the alkoxy group is two methoxy groups), laricitrin formed by the elimination of the methyl group of one methoxy group, and myricetin formed by the elimination of the methyl groups of two methoxy groups. Examples include galangin formed by the elimination of the methyl group of one methoxy group of izalpinin (wherein the alkoxy group is one methoxy group).

[0043] Examples of the dealkylated flavanone include, for example, 8-prenylnaringenin formed by the elimination of the methyl group of one methoxy group of isoxanthohumol (wherein the alkoxy group is one methoxy group), eriodictyol formed by the elimination of the methyl group of one methoxy group of hesperetin (wherein the alkoxy group is one methoxy group), and the like.

[0044] Examples of the dealkylated isoflavone include, for example, 6-hydroxy daidzein formed by the elimination of the methyl group of one methoxy group of glycitein (wherein the alkoxy group is one methoxy group), genistein formed by the elimination of the methyl group of one methoxy group of biochanin (wherein the alkoxy group is one methoxy group), daidzein formed by the elimination of the methyl group of one methoxy group of formononetin (wherein the alkoxy group is one methoxy group), 6-hydroxy genistein formed by the elimination of the methyl group of one methoxy group of tectrigenin (wherein the alkoxy group is one methoxy group), and the like.

[0045] Examples of the dealkylated chalcone include, for example, demethylxanthohumol formed by the elimination of the methyl group of one methoxy group of xanthohumol (wherein the alkoxy group is one methoxy group), and the like.

[0046] Examples of the dealkylated xanthone include, for example, γ-mangostin formed by the elimination of the methyl group of one methoxy group of α-mangostin (wherein the alkoxy group is one methoxy group), α-mangostin formed by the elimination of the methyl group of one methoxy group of β-mangostin (wherein the alkoxy group is two methoxy groups), γ-mangostin formed by the elimination of the methyl groups of two methoxy groups, and the like.

[0047] Examples of the dealkylated simple phenols include for example, 4-acetylresorcinol formed by the elimination of the methyl group of one methoxy group of paeonol (wherein the alkoxy group is one methoxy group), phenol formed by the elimination of the methyl group of one methoxy group of anisole (wherein the alkoxy group is one methoxy group), and the like.

[0048] (a microorganism having the ability to produce a dealkylated polyphenol by eliminating the alkyl group of the alkoxy group of a polyphenol having an alkoxy group in the side chain) The microorganism used in this embodiment and having the ability to produce a dealkylated polyphenol by eliminating the alkyl group of the alkoxy group of a polyphenol having an alkoxy group in the side chain is not particularly limited as long as it is a microorganism belonging to the genus Acetobacterium having the ability to produce a dealkylated polyphenol by eliminating the alkyl group of the alkoxy group of a polyphenol having an alkoxy group in the side chain.

[0049] Regardless of the species and strain of the microorganism, one of them may be used or two or more of them may be used. In addition, the microorganism belonging to the genus Acetobacterium can be read as a bacterium belonging to the genus Acetobacterium.

[0050] Examples of the microorganism belonging to the genus Acetobacterium include, for example, a microorganism belonging to Acetobacterium bakii (such as DSM 8239 strain), a microorganism belonging to Acetobacterium dehalogenans Organisms (e.g., DSM 11527 strain, etc.), microorganisms belonging to Acetobacterium wieringae (e.g., DSM 1911 strain, etc.), Acetobacterium woodii (e.g., DSM 1030 strain, etc.) and the like.

[0051] Taking the Acetobacterium bakii DSM 8239 strain as an example, in this embodiment, the Acetobacterium bakii DSM 8239 strain is not limited to the deposited strain, and may be a strain substantially equivalent to the deposited strain. A strain substantially equivalent to the deposited strain is a strain belonging to the same genus or species as the deposited strain, and having the ability to cleave the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain to produce a dealkylated polyphenol. Also, a strain substantially equivalent to the deposited strain is a strain whose nucleotide sequence of the 16S rRNA gene has a homology of 98.5% or more, preferably 98.7% or more, more preferably 99% or more, and still more preferably 100% with the nucleotide sequence of the 16S rRNA gene of the deposited strain. Further, as long as the deposited strain has the ability to cleave the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain to produce a dealkylated polyphenol, it may be a strain bred from the deposited strain or a strain substantially equivalent thereto by mutation treatment, genetic recombination, selection of natural mutants, etc. This also applies to other deposited strains described above.

[0052] (Resting cells of microorganisms having the ability to cleave the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain to produce a dealkylated polyphenol) In this embodiment, the microorganisms having the ability to cleave the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain to produce a dealkylated polyphenol include their resting cells. The static body refers to a microbial body obtained by removing culture medium components from cultured microorganisms through operations such as centrifugation, washing with a salt solution or buffer solution, and suspending in the same solution as the washing solution, and is a microbial body in a non-growing state. In this embodiment, it refers to a microbial body having at least a metabolic system capable of eliminating the alkyl group of the alkoxy group of a polyphenol having an alkoxy group in the side chain to generate a dealkylated polyphenol. When the microorganism is a bacterium having the ability to eliminate the alkyl group of the alkoxy group of a polyphenol having an alkoxy group in the side chain to generate a dealkylated polyphenol, the static body is a static cell. When the microorganism is a bacterium having the ability to eliminate the alkyl group of the alkoxy group of a polyphenol having an alkoxy group in the side chain to generate a dealkylated polyphenol, the static body is a static cell. Examples of the salt solution include physiological saline. Examples of the buffer solution include phosphate buffer, Tris-hydrochloride buffer, citrate-phosphate buffer, citrate buffer, MOPS buffer, acetate buffer, glycine buffer, etc. In each case, the pH and concentration can be appropriately adjusted according to conventional methods. All the microorganisms of the present disclosure include static bodies.

[0053] (Solution containing polyphenol having an alkoxy group in the side chain) In this embodiment, the solution containing a polyphenol having an alkoxy group in the side chain is not particularly limited as long as the microorganism can eliminate the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain in the solution to generate a dealkylated polyphenol. Preferably it is a culture medium, more preferably the culture medium described in the column of "Culture medium and generation of dealkylated polyphenol by culture" described later. When the microorganism is a static body, the above-mentioned salt solution or buffer solution is preferable. In addition, the "culture medium" in the present disclosure refers to any solution in which microorganisms can grow, including a minimal medium, and does not include solutions in which microorganisms cannot grow, such as the above-mentioned salt solution and buffer solution.

[0054] When adding the polyphenol having an alkoxy group in the side chain to the solution, it may be added before the generation of the dealkylated polyphenol, during the process, and may be added all at once, sequentially, or continuously. The content of the polyphenol having an alkoxy group in the side chain in the solution is usually 0.001 g / L, preferably 0.01 g / L or more, more preferably 0.1 g / L or more, and still more preferably 1 g / L or more. On the other hand, it is usually 100 g / L or less, preferably 20 g / L or less, and more preferably 10 g / L or less.

[0055] (Medium and Generation of Demethylated Polyphenols by Cultivation) In the step (a1), it is preferable that the solution is a medium. The medium is not particularly limited. For example, ANAEROBE BASAL BROTH (ABB medium) manufactured by Oxoid, Wilkins-Chalgren Anaerobe Broth (CM0643) manufactured by Oxoid, GAM medium manufactured by Nissui Pharmaceutical Co., Ltd., modified GAM medium, and Inhart infusion medium and the like can be used.

[0056] In addition, water-soluble organic substances can be added to the medium as a carbon source. Examples of the water-soluble organic substances include the following compounds. That is, sugars such as glucose, arabinose, sorbitol, fructose, mannose, sucrose, trehalose, and xylose; alcohols such as glycerol; and organic acids such as valeric acid, butyric acid, propionic acid, acetic acid, formic acid, and fumaric acid.

[0057] The concentration of the organic substance added to the medium as a carbon source can be appropriately adjusted for efficient growth. Generally, the addition amount can be selected from the range of 0.1 to 10 wt / vol%.

[0058] In addition to the above carbon source, a nitrogen source can be added to the medium. As the nitrogen source, various nitrogen compounds that can be used in ordinary fermentation can be used. Preferred inorganic nitrogen sources include ammonium salts, nitrates, etc., and more preferably ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium hydrogen phosphate, potassium nitrate, and sodium nitrate. In addition, examples of the organic nitrogen source include amino acids, yeast extract, peptones (such as polypeptone N, soy peptone, etc.), meat extract (such as Ehrlich's bonito extract, Lab-Lemco powder, bouillon, etc.), seafood extract, liver extract, digested serum powder, fish oil, etc. It is possible.

[0059] Furthermore, in addition to the carbon source and nitrogen source, for example, by adding cofactors such as vitamins and inorganic compounds such as various salts to the medium, growth and activity may be enhanced in some cases. For example, the following can be cited as microbial growth promoting factors derived from animals and plants, such as inorganic compounds, vitamins, and fatty acids.

[0060] 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 selenate Ammonium ferrous sulfate Sodium acetate trihydrate Magnesium sulfate heptahydrate Manganese sulfate tetrahydrate

[0061] In addition, adding reducing agents such as cysteine, cystine, sodium sulfide, sulfite, ascorbic acid, glutathione, thioglycolic acid, rutin, etc. and enzymes that decompose reactive oxygen species such as catalase and superoxide mutase to the medium may improve growth, and in that case, it is preferable.

[0062] The gas phase and aqueous phase during the culture preferably do not contain air or oxygen, and may, for example, contain nitrogen and / or hydrogen at any ratio, or nitrogen and / or carbon dioxide at any ratio. A gas phase or aqueous phase containing hydrogen is preferred. The ratio of hydrogen in the gas phase is usually 0.5 vol% or more, since this promotes the production of dealkylated polyphenols. The content is preferably 1.0 vol% or more, more preferably 2.0 vol% or more, while it is usually 100 vol% or less, preferably 20 vol% or less, more preferably 10 vol% or less.

[0063] The method for creating such an environment in the gas phase and aqueous phase during culture is not particularly limited, but examples of the method include replacing the gas phase with the gas before culture, supplying the gas from the bottom of the culture vessel and / or to the gas phase of the culture vessel during culture, and bubbling the aqueous phase with the gas before culture. As the hydrogen, hydrogen gas may be used as it is. Alternatively, a source of hydrogen such as formic acid and / or a salt thereof may be added to the culture medium, and hydrogen may be generated during culture by the action of the microorganism.

[0064] The amount of ventilation is preferably 0.005 to 2 vvm, and more preferably 0.05 to 0.5 vvm. The mixed gas can also be supplied in the form of nanobubbles. The culture temperature is preferably 20° C. to 45° C., more preferably 25° C. to 40° C., and even more preferably The temperature is usually between 30℃ and 37℃. The pressurization conditions for the culture vessel are not particularly limited as long as they allow growth, but are preferably in the range of 0.001 to 1 MPa, and more preferably 0.01 to 0.5 MPa. The culture time is preferably 8 to 340 hours, more preferably 12 to 170 hours, and further preferably 16 to 120 hours.

[0065] In addition, it may be preferable to add a surfactant, an adsorbent, an inclusion compound, etc. to the culture solution because the production of dealkylated polyphenols can be promoted in such a case. Examples of the surfactant include Tween 80, etc., and it can be added in an amount of about 0.001 g / L or more and 10 g / L or less. Examples of the adsorbent include cellulose and its derivatives; dextrin; Diaion HP series and Sepabeads series which are hydrophobic adsorbents manufactured by Mitsubishi Chemical Corporation; Amberlite XAD series manufactured by Organo Corporation, etc.

[0066] Examples of the inclusion compound include α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, cluster dextrin (highly branched cyclic dextrin), and in addition, their analogs may also be used, for example, methyl-β-cyclodextrin, trimethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, etc. Among these, γ-cyclodextrin may be the most effective, and in such a case, it is preferable. In addition, since the production of dealkylated polyphenols can sometimes be further promoted by coexisting two or more kinds of inclusion compounds, it is preferable in such a case. The addition amount is the total amount of the molar ratio when the total amount of the polyphenol having an alkoxy group in the side chain is taken as 1, usually 0.1 or more, preferably 0.5 or more, more preferably 1.0 or more, while usually 5.0 or less, preferably 2.5 or less, more preferably 2.0 or less.

[0067] (Production of dealkylated polyphenols by resting cells) When the microorganism is a resting cell, the solution is preferably a salt solution or a buffer solution described in the column of "Resting cells of a microorganism having the ability to produce dealkylated polyphenols by eliminating the alkyl group of the alkoxy group of the polyphenol having an alkoxy group in the side chain" instead of the above-mentioned medium. Regarding other conditions, they are the same as the description in the column of "Medium and production of dealkylated polyphenols by culture".

[0068] (Other processes) This embodiment may include, for example, a step of quantifying the obtained dealkylated polyphenol. The method can follow a conventional method. For example, a part of the culture solution is collected, appropriately diluted, stirred well, filtered using a membrane such as a polytetrafluoroethylene (PTFE) membrane to remove insolubles, and the resulting solution is quantified by high performance liquid chromatography, etc.

[0069] Also, this embodiment may include a step of recovering the obtained dealkylated polyphenol. The recovery step includes a purification step, a concentration step, etc. As the purification treatment in the purification step, sterilization of microorganisms by heat, etc.; sterilization by microfiltration (MF), ultrafiltration (UF), etc.; removal of solids and macromolecular substances; extraction with an organic solvent or an ionic liquid, etc.; adsorption and decolorization using a hydrophobic adsorbent, an ion exchange resin, an activated carbon column, etc. can be performed. Also, as the concentration treatment in the concentration step, concentration by an evaporator, a reverse osmosis membrane, etc. can be mentioned. Furthermore, the solution containing the obtained dealkylated polyphenol can be powdered by freeze drying, spray drying, etc. In powdering, excipients such as lactose, dextrin, corn starch, etc. can also be added.

[0070] <Method for producing equol> Another aspect of the present disclosure includes the above step (a1) and the following step (a2) performed in the same system, wherein the polyphenol having an alkoxy group in the side chain in the step (a1) is glycitein, and the dealkylated polyphenol is 6-hydroxy daidzein, which is a method for producing equol. Step (a2): A step of causing a microorganism having the ability to produce equol from 6-hydroxy daidzein to produce equol from 6-hydroxy daidzein in the solution containing 6-hydroxy daidzein generated in the step (a1)

[0071] (Microorganisms having the ability to produce equol from 6-hydroxy daidzein) In the solution containing the above 6-hydroxy daidzein, the microorganism having the ability to produce equol from 6-hydroxy daidzein is not particularly limited as long as it is a microorganism having the ability to produce equol from 6-hydroxy daidzein in the solution containing 6-hydroxy daidzein. The above microorganism can be obtained by ordinary screening methods. For example, when culturing a microorganism in a solution containing 6-hydroxy daidzein according to an ordinary culturing method, a microorganism capable of producing equol from 6-hydroxy daidzein may be selected. The above microorganism is preferably a bacterium. Examples of the above bacterium include intestinal bacteria.

[0072] Examples of the above intestinal bacteria include microorganisms belonging to the genus Adlercreutzia microorganisms belonging to the genus Lactoccocus, microorganisms belonging to the genus Eggerthella, microorganisms belonging to the genus Asaccharobacter, etc. 。 Regardless of the species and strain of the above microorganism, one of them may be used or two or more of them may be used. In addition, the microorganism belonging to the genus Adlercreutzia can be read as a bacterium belonging to the genus Adlercreutzia, the microorganism belonging to the genus Lactoccocus can be read as a bacterium belonging to the genus Lactoccocus, the microorganism belonging to the genus Eggerthella can be read as a bacterium belonging to the genus Eggerthella, and the microorganism belonging to the genus Asaccharobacter can be read as a bacterium belonging to the genus Asaccharobacter.

[0073] Examples of the microorganism belonging to the genus Adlercreutzia include, for example, Adler a microorganism belonging to Adlercreutzia equolifaciens ([[]] for example, the DSM 19450 strain, etc.).

[0074] Examples of the microorganism belonging to the genus Lactoccocus include, for example, Lactocc a microorganism belonging to Lactococcus garvieae, etc.

[0075] Examples of the microorganism belonging to the genus Eggerthella include, for example, Eggerthella sp YY 7918 strain, etc.

[0076] Examples of the microorganism belonging to the genus Asaccharobacter include, for example, A a microorganism belonging to Asaccharobater celatus (for example, the DSM 18785 strain, etc.).

[0077] Although the details of the above microorganisms are as described above, taking the Adlercreutzia equolifaciens DSM 19450 strain as an example, in this embodiment, Adler The strain Adlercreutzia equolifaciens DSM 19450 is not limited to the deposited strain, and may be a strain substantially equivalent to the deposited strain. A strain substantially equivalent refers to a strain belonging to the same genus or species as the deposited strain and having the ability to produce equol from 6-hydroxy daidzein. Also, a strain substantially equivalent refers to a strain whose nucleotide sequence of the 16S rRNA gene has a homology of 98.5% or more, preferably 98.7% or more, more preferably 99% or more, and even more preferably 100% with the nucleotide sequence of the 16S rRNA gene of the deposited strain. Further, as long as the deposited strain has the ability to produce equol from 6-hydroxy daidzein, it may be a strain bred from the deposited strain or a strain substantially equivalent thereto by mutation treatment, genetic recombination, selection of natural mutants, etc. This also applies equally to other deposited strains already described.

[0078] (Resting cells of microorganisms that produce equol from 6-hydroxy daidzein) Microorganisms that produce equol from 6-hydroxy daidzein include their resting cells. Regarding the resting cells, it is the same as the description in the column of "Resting cells of microorganisms having the ability to eliminate the alkyl group of the alkoxy group in the side chain of polyphenols having an alkoxy group to produce dealkylated polyphenols".

[0079] (Solution containing 6-hydroxy daidzein) A solution containing 6-hydroxy daidzein is a solution containing 6-hydroxy daidzein, which is the dealkylated polyphenol produced in the step (a1).

[0080] Separate from the 6-hydroxy daidzein produced in the step (a1), when further 6-hydroxy daidzein is added to the solution, it may be added before the production of equol, during the process, and may be added all at once, sequentially, or continuously. The content of 6-hydroxy daidzein in the solution is usually 0.01 g / L or more, preferably 0.1 g / L or more, more preferably 1 g / L or more. On the other hand, it is usually 100 g / L or less, preferably 20 g / L or less, more preferably 10 g / L or less.

[0081] (Medium and production of equol by culture) In the method for producing equol according to this aspect, the steps (a1) and (a2) are carried out in the same system. Therefore, the medium conditions in step (a2) and the conditions for producing equol by culture are the same as those described in the section of "Medium and production of dealkylated polyphenols by culture".

[0082] (Production of equol by resting cells) In the method for producing equol according to this aspect, the steps (a1) and (a2) are carried out in the same system. Therefore, when the microorganism that produces equol from 6-hydroxy daidzein is a resting cell, the solution is also preferably a salt solution or buffer solution described in the section of "Resting cells of microorganisms having the ability to produce dealkylated polyphenols by eliminating the alkyl group of the alkoxy group of polyphenols having an alkoxy group in the side chain". Regarding other conditions, they are the same as those described in the section of "Medium and production of dealkylated polyphenols by culture".

[0083] (Same system) That the steps (a1) and (a2) are carried out in the same system means that a series of processes from the generation of 6-hydroxy daidzein in step (a1) until the generated 6-hydroxy daidzein is directly used as the 6-hydroxy daidzein in step (a2) and equol is generated in step (a2) are continuously carried out in the same system. That is, between step (a1) and step (a2), for example, there is no step of separating and / or purifying the 6-hydroxy daidzein generated in step (a1). It means that it does not include steps such as separating and / or purifying the 6-hydroxy daidzein generated in step (a1).

[0084] As a specific example, inoculating and culturing a microorganism having the ability to produce 6-hydroxy daidzein from glycitein and a microorganism having the ability to produce equol from 6-hydroxy daidzein in the same medium can produce equol.

[0085] (Other steps) This embodiment may include, for example, a step of quantifying the obtained equol or a step of recovering the obtained equol. The details are the same as the description in the "Other steps" section of the "Method for producing dealkylated polyphenols". Furthermore, the solution containing equol can be powdered by freeze-drying, spray-drying, etc. In powdering, excipients such as lactose, dextrin, and corn starch can also be added.

[0086] Examples are described below, but none of the examples are to be construed as limiting examples.

[0087] [Example 1] After adding isoxanthohumol (manufactured by Nacalai Tesque) to ABB medium (manufactured by Oxoid), it was heat-sterilized, and the gas phase was replaced with N 2 :CO 2 :H 2 (80% / 10% / 10%) gas to obtain a basic medium. Acetobacterium bakii DSM 8239 strain was inoculated into the medium containing 5 mg / L of isoxanthohumol at a final concentration and cultured anaerobically at 37°C. After the culture was completed, prenylflavonoids were extracted from 5 mL of the culture solution with an equal volume of ethyl acetate (1.5% formic acid). After recovering the obtained ethyl acetate phase, it was dried. The dried product thus obtained was redissolved in 0.5 mL of methanol, and quantitative analysis of prenylflavonoids was performed by HPLC. HPLC was performed under the conditions described below. Prenylflavonoids manufactured by LKT Laboratories were used as a standard and dissolved in DMSO for use.

[0088] <HPLC Conditions> Column: Inertsil ODS-3 (250×4.6 mm) (manufactured by GL Science) Eluent: Solution A (water / formic acid = 99 / 1), Solution B (acetonitrile / formic acid = 99 / 1), and a gradient of 20% - 70% of Solution B Flow rate: 1.0 mL / min Column temperature: 40 °C Detection: 290 nm

[0089] The results are shown in Table 1. After culturing for one week, 1.35% of the substrate isoxanthohumol was converted to 8-prenylnaringenin.

[0090]

Table 1

[0091] [Example 2] 5 mL of the modified GAM medium manufactured by Nissui Pharmaceutical Co., Ltd. was dispensed into a test tube. Then, it was gas-substituted with nitrogen gas and sterilized. This was used as the pre-culture medium. Glycitein was added to the modified GAM medium to a concentration of 54 mg / L, and 10 mL was dispensed into a test tube. Then, it was gas-substituted with nitrogen gas and sterilized. This was used as the main-culture medium. After inoculating the pre-culture medium with Acetobacterium bakii DSM 8239 strain, it was gas-substituted with nitrogen gas and cultured at 37 °C and 200 spm for 1 day (pre-culture). After inoculating the above pre-cultured strain into the main-culture medium, it was gas-substituted with nitrogen gas and cultured at 37 °C and 200 spm for 7 days (main-culture).

[0092] 20 μL was sampled from the main-culture medium after culturing, diluted 50-fold with a diluent (ethanol: MilliQ water = 70:30 (v / v)). After filtering this through a 0.45 μm filter, the supernatant was analyzed under the following HPLC conditions.

[0093] <HPLC Conditions> Column: Phenomenex SYNERGI 4μm POLAR-R 150mm×4.6mm Eluent: Distilled water / methanol = 55 / 45 (v / v) Temperature: 40 °C Detection: 280 nm Flow rate: 1.0 mL / min Injection volume: 10 μL Time: 30 min

[0094] The results are shown in Table 2. After 7 days of culture, 37.6% of the substrate glycitein was converted to 6-hydroxy daidzein.

[0095]

Table 2

[0096] [Reference Example 1] 5 mL of the modified GAM medium manufactured by Nissui Pharmaceutical Co., Ltd. was dispensed into a test tube. Then, it was gas-substituted with nitrogen gas and sterilized. This was used as the preculture medium. 6-Hydroxy daidzein was added to the modified GAM medium to a concentration of 50 mg / L, and 10 mL was dispensed into a test tube. Then, it was gas-substituted with nitrogen gas and sterilized. This was used as the main culture medium. Asaccharobater celatus DSM 18785 was inoculated into the preculture medium, then gas-substituted with nitrogen gas, and cultured at 37 °C and 200 spm for 1 day (preculture). . After inoculating the above precultured strain into the main culture medium, it was gas-substituted with nitrogen gas and cultured at 37 °C and 200 spm for 3 days (main culture). 20 μL was sampled from the main culture medium after culture and diluted 50-fold with a diluent (ethanol: Milli-Q water = 70:30 (v / v)). After filtering this through a 0.45 μm filter, the supernatant was analyzed under the following HPLC conditions.

[0097] <HPLC Conditions> Column: Phenomenex SYNERGI 4μm POLAR-R 150mm×4.6mm Eluent: Distilled water / methanol = 55 / 45 (v / v) Temperature: 40 °C Detection: 280 nm Flow rate: 1.0 mL / min Injection volume: 10 μL Time: 30 min

[0098] The results are shown in Table 3. By culturing for 3 days, 48.8% of the substrate 6-hydroxy daidzein was converted to equol.

[0099]

Table 3

Industrial Applicability

[0100] The dealkylated polyphenol produced by the production method according to one aspect of the present disclosure is useful as a raw material or material for cosmetics, quasi-drugs, medical supplies, sanitary products, pharmaceuticals, food and beverages (including supplements), etc.

Claims

1. A method for producing 8-prenylnaringenin, comprising the following steps. Step: In a solution containing isoxanthohumol, allowing a microorganism belonging to the genus Acetobacterium bakii, which has the ability to produce 8-prenylnaringenin from isoxanthohumol, to produce 8-prenylnaringenin from isoxanthohumol

2. A method for producing 6-hydroxy daidzein, comprising the following steps. Step: In a solution containing glycitein, allowing a microorganism belonging to the genus Acetobacterium bakii, which has the ability to produce 6-hydroxy daidzein from glycitein, to produce 6-hydroxy daidzein from glycitein

3. The production method according to claim 1 or 2, wherein the microorganism belonging to the genus Acetobacterium bakii is Acetobacterium bakii DSM 8239 strain.

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