Biotransformation of C-Glucosylisoflavone Puerarin to Estrogenic (3S)-Equol in Co-culture of Two Human Intestinal Bacteria

Jin, Jong-Sik; Nishihata, Tomohiro; Kakiuchi, Nobuko; Hattori, Masao

doi:10.1248/bpb.31.1621

Cited by 78 publications

(49 citation statements)

References 27 publications

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“…Lower values of sequence identity were observed for strain HE8 and the equol-forming species Adlercreutzia equolifaciens (90.2%), Eggerthella sp. strain Julong 732 (89.9%) and strain DZE (87.6%) from human feces, Asaccharobacter celatus (90.1%) from rat cecum, and strain Mt1B8 from mouse intestine (88.8%), all of which affiliate within the Coriobacteriaceae (13,16,17,20,21,36). All of these bacterial strains produce equol from daidzein, with the exception of Eggerthella sp.…”

Section: Vol 75 2009 Daidzein and Genistein Conversion 1741mentioning

confidence: 99%

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Isolation of a Human Intestinal Bacterium Capable of Daidzein and Genistein Conversion

Matthies

Blaut

Braune

2009

Appl Environ Microbiol

147

123

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Section: Vol 75 2009 Daidzein and Genistein Conversion 1741mentioning

confidence: 99%

“…While formation of equol from daidzein via dihydrodaidzein is catalyzed by several bacterial species (16,17,20,21,40), the corresponding conversion of genistein to 5-hydroxy-equol has been demonstrated for only two strains, Mt1B8 and DZE (13,17). This may be due to the fact that genistein transformation has not been investigated.…”

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confidence: 99%

Isolation of a Human Intestinal Bacterium Capable of Daidzein and Genistein Conversion

Matthies

Blaut

Braune

2009

Appl Environ Microbiol

147

123

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“…Daidzein is thought to be metabolized by human intestinal bacteria to equol or to Odesmethylangolensin via dihydrodaidzein and tetrahydrodaidzein (14,15,22,32); however, neither the enzymes involved in the metabolism of daidzein to equol nor even the metabolic pathway has been clarified fully for equol-producing bacteria.…”

mentioning

confidence: 99%

Cloning and Expression of a Novel NADP(H)-Dependent Daidzein Reductase, an Enzyme Involved in the Metabolism of Daidzein, from Equol-Producing Lactococcus Strain 20-92

Shimada

Yasuda

Takahashi

et al. 2010

Appl Environ Microbiol

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Equol is a metabolite produced from daidzein by enteric microflora, and it has attracted a great deal of attention because of its protective or ameliorative ability against several sex hormone-dependent diseases (e.g., menopausal disorder and lower bone density), which is more potent than that of other isoflavonoids. We It was further suggested that L-DZNR was an NAD(H)/NADP(H):flavin oxidoreductase belonging to the old yellow enzyme (OYE) family. Recombinant histidine-tagged L-DZNR was expressed in Escherichia coli.The recombinant protein converted daidzein to (S)-dihydrodaidzein with enantioselectivity. This is the first report of the isolation of an enzyme related to daidzein metabolism and equol production in enteric bacteria.Isoflavones are flavonoids present in various plants and are known to be abundant in soybeans and legumes. These compounds have been called phytoestrogens because their chemical structure is similar to that of the female sex hormone, estrogen. Isoflavones have an ability to bind to estrogen receptors and show protection against or improvement in several sex hormone-dependent diseases, such as breast cancer, prostate cancer, menopausal disorder, lower bone density, and hypertension, due to their weak agonistic or antagonistic effects (1,19,27).Daidzein is one of the main soy isoflavonoids produced from daidzin by the glucosidase of intestinal bacteria (17). Equol is a metabolite produced from daidzein by the enterobacterial microflora (5). Recently, equol has attracted a great deal of attention because its estrogenic activity is more potent than that of other isoflavonoids, including daidzein (27). It is well known that individual variation exists in the ability of these enteric microflora to produce equol and that less than half the human population is capable of producing equol after ingesting soy isoflavones (3). Therefore, to increase the production of equol in the enteric environment of each individual, the development of probiotics using safe bacteria which have the ability to produce equol from daidzein is ongoing.Lactococcus strain 20-92 (Lactococcus 20-92; 30a) is an equol-producing lactic acid bacterium isolated from the feces of healthy humans by Uchiyama et al. (30). This bacterium is spherical and Gram positive and is a strain of L. garvieae. The application of Lactococcus 20-92 in probiotics is advantageous because L. garvieae is not pathogenic or toxic to humans.To date, other bacterial strains that are capable of transforming daidzein to dihydrodaidzein or equol have been isolated (9,21,22,23,29,32,36,37). Daidzein is thought to be metabolized by human intestinal bacteria to equol or to Odesmethylangolensin via dihydrodaidzein and tetrahydrodaidzein (14,15,22,32); however, neither the enzymes involved in the metabolism of daidzein to equol nor even the metabolic pathway has been clarified fully for equol-producing bacteria.In this study, we purified an enzyme from Lactococcus 20-92 that assisted in the conversion of daidzein to dihydrodaidzein. Furthermore, we cloned the...

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“…1) Puerarin was isolated from the roots of Pueraria lobata (WILLD.) OHWI (Tochimoto Tenkaido Co., Osaka, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…1,[3][4][5][6][7][8][9][10][11] Despite of the growing importance of C-glycosides in the field of pharmaceuticals, characteristic features of C-glucosyl bond-cleaving enzymes involved in its metabolism were still remain to be unclear.…”

mentioning

confidence: 99%

The C-Glucosyl Bond of Puerarin Was Cleaved Hydrolytically by a Human Intestinal Bacterium Strain PUE to Yield Its Aglycone Daidzein and an Intact Glucose

Nakamura

Nishihata

Jin

et al. 2011

CHEMICAL & PHARMACEUTICAL BULLETIN

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Many types of glycosides are widely distributed as major constituents in medicinal plants. However, due to their hydrophilic properties, they are poorly absorbed from gastro intestinal tract into body fluid, but their aglycones formed by intestinal bacterial hydrolysis are readily absorbed. Therefore, intestinal bacteria play an important role in increasing absorption and efficacy of drugs including glycosides. In nature, there is a special type of glycoside, C-glycosides, such as those of flavone, isoflavone, chromone, xanthone, anthrone, and gallic acid. Since C-1 of the sugar ring is directly connected to the aglycone by C-C bonding, C-glycosides are usually resistant against acidic and enzymatic hydrolysis in contrast with the corresponding O-glycosides. Thus, with the prospect of improving the stability, various C-glycoside analogues mimicking bioactive but labile O-glycosides have been prepared for developing the new drugs.2) In addition, a lot of publications describing in detail about O-glycosidase appeared, but that of C-glycosidase did not.On the other aspects, in the last two decades, C-glucosyl bond-cleaving reactions by human intestinal bacteria were reported. 1,[3][4][5][6][7][8][9][10][11] Despite of the growing importance of C-glycosides in the field of pharmaceuticals, characteristic features of C-glucosyl bond-cleaving enzymes involved in its metabolism were still remain to be unclear.Recently, we isolated a new bacterial species named strain PUE that transforms a C-glucoside puerarin to its aglycone daidzein.1) The aglycone moiety is normally detected easily because of UV active, however, the liberated sugar segment from C-glucoside has not been yet identified. The structural elucidation of the liberated sugar moiety could yield a valuable clue in revealing the reaction mechanism (Fig. 1). In the present paper, we describe the structural elucidation of the sugar moiety after cleavage of a C-glucosyl bond of puerarin by anaerobic incubation with a human intestinal bacterium, strain PUE, and a cell-free extract. Results Synthesis of [6؆,6؆-D 2 ]puerarin (8)Since puerarin (1) has only one primary alcohol at C-6 of the sugar moiety, we targeted it for labeling with deuterium. First, 1 was treated with benzaldehyde and zinc chloride to give 2. Other hydroxyl groups of 2 were acetylated with acetic anhydride in pyridine to obtain 3. Benzylidene acetal of 3 was hydrolyzed under an acidic condition to afford 4. The primary hydroxyl of 4 was oxidized to carboxylic acid with a Jones reagent to give 5. The acetyl groups of 5 were removed by alkali to give 6, then, the carboxyl group was esterified to yield 7 by a usual method. We failed the reduction of 7 to the corresponding alcohol with LiAlD 4 due to decomposition, but 8 was prepared from 7 by slow addition of CD 3 OD into the reaction mixture containing NaBD 4 . 12) In this reaction, the addition of CH 3 OH instead of CD 3 OD gave a mixture of 8 and [6Љ-H,6Љ-D]puerarin (ca. 1 : 1 ratio), because hydrogen atoms originated from CH 3 OH were also incorp...

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Biotransformation of C-Glucosylisoflavone Puerarin to Estrogenic (3S)-Equol in Co-culture of Two Human Intestinal Bacteria

Cited by 78 publications

References 27 publications

Isolation of a Human Intestinal Bacterium Capable of Daidzein and Genistein Conversion

Isolation of a Human Intestinal Bacterium Capable of Daidzein and Genistein Conversion

Cloning and Expression of a Novel NADP(H)-Dependent Daidzein Reductase, an Enzyme Involved in the Metabolism of Daidzein, from Equol-Producing Lactococcus Strain 20-92

The C-Glucosyl Bond of Puerarin Was Cleaved Hydrolytically by a Human Intestinal Bacterium Strain PUE to Yield Its Aglycone Daidzein and an Intact Glucose

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