Daidzein reductase of Eggerthella sp. YY7918, its octameric subunit structure containing FMN/FAD/4Fe-4S, and its enantioselective production of R-dihydroisoflavones
S-Equol is a metabolite of daidzein, a type of soy isoflavone, and three reductases are involved in the conversion of daidzein by specific intestinal bacteria. S-Equol is thought to prevent hormone-dependent diseases. We previously identified the equol producing gene cluster (eqlABC) of Eggerthella...
Gespeichert in:
Veröffentlicht in: | Journal of bioscience and bioengineering 2018-09, Vol.126 (3), p.301-309 |
---|---|
Hauptverfasser: | , , , , , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 309 |
---|---|
container_issue | 3 |
container_start_page | 301 |
container_title | Journal of bioscience and bioengineering |
container_volume | 126 |
creator | Kawada, Yuika Goshima, Tomoko Sawamura, Rie Yokoyama, Shin-ichiro Yanase, Emiko Niwa, Toshio Ebihara, Akio Inagaki, Mizuho Yamaguchi, Keiichi Kuwata, Kazuo Kato, Yuta Sakurada, Osamu Suzuki, Tohru |
description | S-Equol is a metabolite of daidzein, a type of soy isoflavone, and three reductases are involved in the conversion of daidzein by specific intestinal bacteria. S-Equol is thought to prevent hormone-dependent diseases. We previously identified the equol producing gene cluster (eqlABC) of Eggerthella sp. YY7918. Daidzein reductase (DZNR), encoded by eqlA, catalyzes the reduction of daidzein to dihydrodaidzein (the first step of equol synthesis), which was confirmed using a recombinant enzyme produced in Escherichia coli. Here, we purified recombinant DZNR to homogeneity and analyzed its enzymological properties. DZNR contained FMN, FAD, and one 4Fe-4S cluster per 70-kDa subunit as enzymatic cofactors. DZNR reduced the CC bond between the C-2 and C-3 positions of daidzein, genistein, glycitein, and formononetin in the presence of NADPH. R-Dihydrodaidzein and R-dihydrogenistein were highly stereo-selectively produced from daidzein and genistein. The Km and kcat for daidzein were 11.9 μM and 6.7 s−1, and these values for genistein were 74.1 μM and 28.3 s−1, respectively. This enzyme showed similar kinetic parameters and wide substrate specificity for isoflavone molecules. Thus, this enzyme appears to be an isoflavone reductase. Gel filtration chromatography and chemical cross-linking analysis of the active form of DZNR suggested that the enzyme consists of an octameric subunit structure. We confirmed this by small-angle X-ray scattering and transmission electron microscopy at a magnification of ×200,000. DZNR formed a globular four-petal cloverleaf structure with a central vertical hole. The maximum particle size was 173 Å. |
doi_str_mv | 10.1016/j.jbiosc.2018.03.018 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2032433027</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S1389172317309817</els_id><sourcerecordid>2032433027</sourcerecordid><originalsourceid>FETCH-LOGICAL-c492t-f26cc842c29e1df6b54e3ef6cb81eefe4b57675d71ba603920c4526b4140fe963</originalsourceid><addsrcrecordid>eNp9kc1u1DAUhS0EoqXlDRDykkWT8d8k8QapajuA1IJEYdGV5dg3U48y9tR2RirP0wfFYQpLVseL7_jcew9C7yipKaHNYlNveheSqRmhXU14XeQFOqZctJUQjL6c352saMv4EXqT0oYQ2pKWvkZHTDZSSsGO0dOldvYXOI8j2MlknQCHAV-t1xDzPYyjxmlX47u7VtLuDLuccCjUFqIzOE395F3GKcdinSJgE3zWzju_xqubr4vV-eVCrKASt2dYe_vHDl77XAaHEUx2e8C7GOZkF_yc_L2y7v7RxuBSGEa9Dx7SKXo16DHB22c9QT9XVz8uPlfX3z59uTi_royQLFcDa4zpBDNMArVD0y8FcBga03cUYADRL9umXdqW9rohXDJixJI1vaCCDCAbfoI-HP4tIz1MkLLaumTmI3gIU1KMcCY4J6wtqDigJoaUIgxqF91Wx0dFiZr7URt16EfN_SjCVZFie_-cMPVbsP9MfwspwMcDAGXPvYOoknHgDVgXy72UDe7_Cb8B2uWlNw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2032433027</pqid></control><display><type>article</type><title>Daidzein reductase of Eggerthella sp. YY7918, its octameric subunit structure containing FMN/FAD/4Fe-4S, and its enantioselective production of R-dihydroisoflavones</title><source>Access via ScienceDirect (Elsevier)</source><creator>Kawada, Yuika ; Goshima, Tomoko ; Sawamura, Rie ; Yokoyama, Shin-ichiro ; Yanase, Emiko ; Niwa, Toshio ; Ebihara, Akio ; Inagaki, Mizuho ; Yamaguchi, Keiichi ; Kuwata, Kazuo ; Kato, Yuta ; Sakurada, Osamu ; Suzuki, Tohru</creator><creatorcontrib>Kawada, Yuika ; Goshima, Tomoko ; Sawamura, Rie ; Yokoyama, Shin-ichiro ; Yanase, Emiko ; Niwa, Toshio ; Ebihara, Akio ; Inagaki, Mizuho ; Yamaguchi, Keiichi ; Kuwata, Kazuo ; Kato, Yuta ; Sakurada, Osamu ; Suzuki, Tohru</creatorcontrib><description>S-Equol is a metabolite of daidzein, a type of soy isoflavone, and three reductases are involved in the conversion of daidzein by specific intestinal bacteria. S-Equol is thought to prevent hormone-dependent diseases. We previously identified the equol producing gene cluster (eqlABC) of Eggerthella sp. YY7918. Daidzein reductase (DZNR), encoded by eqlA, catalyzes the reduction of daidzein to dihydrodaidzein (the first step of equol synthesis), which was confirmed using a recombinant enzyme produced in Escherichia coli. Here, we purified recombinant DZNR to homogeneity and analyzed its enzymological properties. DZNR contained FMN, FAD, and one 4Fe-4S cluster per 70-kDa subunit as enzymatic cofactors. DZNR reduced the CC bond between the C-2 and C-3 positions of daidzein, genistein, glycitein, and formononetin in the presence of NADPH. R-Dihydrodaidzein and R-dihydrogenistein were highly stereo-selectively produced from daidzein and genistein. The Km and kcat for daidzein were 11.9 μM and 6.7 s−1, and these values for genistein were 74.1 μM and 28.3 s−1, respectively. This enzyme showed similar kinetic parameters and wide substrate specificity for isoflavone molecules. Thus, this enzyme appears to be an isoflavone reductase. Gel filtration chromatography and chemical cross-linking analysis of the active form of DZNR suggested that the enzyme consists of an octameric subunit structure. We confirmed this by small-angle X-ray scattering and transmission electron microscopy at a magnification of ×200,000. DZNR formed a globular four-petal cloverleaf structure with a central vertical hole. The maximum particle size was 173 Å.</description><identifier>ISSN: 1389-1723</identifier><identifier>EISSN: 1347-4421</identifier><identifier>DOI: 10.1016/j.jbiosc.2018.03.018</identifier><identifier>PMID: 29699942</identifier><language>eng</language><publisher>Japan: Elsevier B.V</publisher><subject>Equol ; FAD ; FMN ; Gut microflora ; Isoflavone ; NADPH ; Old yellow enzyme ; Reductase ; Small-angle X-ray scattering</subject><ispartof>Journal of bioscience and bioengineering, 2018-09, Vol.126 (3), p.301-309</ispartof><rights>2018 The Society for Biotechnology, Japan</rights><rights>Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c492t-f26cc842c29e1df6b54e3ef6cb81eefe4b57675d71ba603920c4526b4140fe963</citedby><cites>FETCH-LOGICAL-c492t-f26cc842c29e1df6b54e3ef6cb81eefe4b57675d71ba603920c4526b4140fe963</cites><orcidid>0000-0002-1763-9084</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jbiosc.2018.03.018$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29699942$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kawada, Yuika</creatorcontrib><creatorcontrib>Goshima, Tomoko</creatorcontrib><creatorcontrib>Sawamura, Rie</creatorcontrib><creatorcontrib>Yokoyama, Shin-ichiro</creatorcontrib><creatorcontrib>Yanase, Emiko</creatorcontrib><creatorcontrib>Niwa, Toshio</creatorcontrib><creatorcontrib>Ebihara, Akio</creatorcontrib><creatorcontrib>Inagaki, Mizuho</creatorcontrib><creatorcontrib>Yamaguchi, Keiichi</creatorcontrib><creatorcontrib>Kuwata, Kazuo</creatorcontrib><creatorcontrib>Kato, Yuta</creatorcontrib><creatorcontrib>Sakurada, Osamu</creatorcontrib><creatorcontrib>Suzuki, Tohru</creatorcontrib><title>Daidzein reductase of Eggerthella sp. YY7918, its octameric subunit structure containing FMN/FAD/4Fe-4S, and its enantioselective production of R-dihydroisoflavones</title><title>Journal of bioscience and bioengineering</title><addtitle>J Biosci Bioeng</addtitle><description>S-Equol is a metabolite of daidzein, a type of soy isoflavone, and three reductases are involved in the conversion of daidzein by specific intestinal bacteria. S-Equol is thought to prevent hormone-dependent diseases. We previously identified the equol producing gene cluster (eqlABC) of Eggerthella sp. YY7918. Daidzein reductase (DZNR), encoded by eqlA, catalyzes the reduction of daidzein to dihydrodaidzein (the first step of equol synthesis), which was confirmed using a recombinant enzyme produced in Escherichia coli. Here, we purified recombinant DZNR to homogeneity and analyzed its enzymological properties. DZNR contained FMN, FAD, and one 4Fe-4S cluster per 70-kDa subunit as enzymatic cofactors. DZNR reduced the CC bond between the C-2 and C-3 positions of daidzein, genistein, glycitein, and formononetin in the presence of NADPH. R-Dihydrodaidzein and R-dihydrogenistein were highly stereo-selectively produced from daidzein and genistein. The Km and kcat for daidzein were 11.9 μM and 6.7 s−1, and these values for genistein were 74.1 μM and 28.3 s−1, respectively. This enzyme showed similar kinetic parameters and wide substrate specificity for isoflavone molecules. Thus, this enzyme appears to be an isoflavone reductase. Gel filtration chromatography and chemical cross-linking analysis of the active form of DZNR suggested that the enzyme consists of an octameric subunit structure. We confirmed this by small-angle X-ray scattering and transmission electron microscopy at a magnification of ×200,000. DZNR formed a globular four-petal cloverleaf structure with a central vertical hole. The maximum particle size was 173 Å.</description><subject>Equol</subject><subject>FAD</subject><subject>FMN</subject><subject>Gut microflora</subject><subject>Isoflavone</subject><subject>NADPH</subject><subject>Old yellow enzyme</subject><subject>Reductase</subject><subject>Small-angle X-ray scattering</subject><issn>1389-1723</issn><issn>1347-4421</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNp9kc1u1DAUhS0EoqXlDRDykkWT8d8k8QapajuA1IJEYdGV5dg3U48y9tR2RirP0wfFYQpLVseL7_jcew9C7yipKaHNYlNveheSqRmhXU14XeQFOqZctJUQjL6c352saMv4EXqT0oYQ2pKWvkZHTDZSSsGO0dOldvYXOI8j2MlknQCHAV-t1xDzPYyjxmlX47u7VtLuDLuccCjUFqIzOE395F3GKcdinSJgE3zWzju_xqubr4vV-eVCrKASt2dYe_vHDl77XAaHEUx2e8C7GOZkF_yc_L2y7v7RxuBSGEa9Dx7SKXo16DHB22c9QT9XVz8uPlfX3z59uTi_royQLFcDa4zpBDNMArVD0y8FcBga03cUYADRL9umXdqW9rohXDJixJI1vaCCDCAbfoI-HP4tIz1MkLLaumTmI3gIU1KMcCY4J6wtqDigJoaUIgxqF91Wx0dFiZr7URt16EfN_SjCVZFie_-cMPVbsP9MfwspwMcDAGXPvYOoknHgDVgXy72UDe7_Cb8B2uWlNw</recordid><startdate>20180901</startdate><enddate>20180901</enddate><creator>Kawada, Yuika</creator><creator>Goshima, Tomoko</creator><creator>Sawamura, Rie</creator><creator>Yokoyama, Shin-ichiro</creator><creator>Yanase, Emiko</creator><creator>Niwa, Toshio</creator><creator>Ebihara, Akio</creator><creator>Inagaki, Mizuho</creator><creator>Yamaguchi, Keiichi</creator><creator>Kuwata, Kazuo</creator><creator>Kato, Yuta</creator><creator>Sakurada, Osamu</creator><creator>Suzuki, Tohru</creator><general>Elsevier B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-1763-9084</orcidid></search><sort><creationdate>20180901</creationdate><title>Daidzein reductase of Eggerthella sp. YY7918, its octameric subunit structure containing FMN/FAD/4Fe-4S, and its enantioselective production of R-dihydroisoflavones</title><author>Kawada, Yuika ; Goshima, Tomoko ; Sawamura, Rie ; Yokoyama, Shin-ichiro ; Yanase, Emiko ; Niwa, Toshio ; Ebihara, Akio ; Inagaki, Mizuho ; Yamaguchi, Keiichi ; Kuwata, Kazuo ; Kato, Yuta ; Sakurada, Osamu ; Suzuki, Tohru</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c492t-f26cc842c29e1df6b54e3ef6cb81eefe4b57675d71ba603920c4526b4140fe963</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Equol</topic><topic>FAD</topic><topic>FMN</topic><topic>Gut microflora</topic><topic>Isoflavone</topic><topic>NADPH</topic><topic>Old yellow enzyme</topic><topic>Reductase</topic><topic>Small-angle X-ray scattering</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kawada, Yuika</creatorcontrib><creatorcontrib>Goshima, Tomoko</creatorcontrib><creatorcontrib>Sawamura, Rie</creatorcontrib><creatorcontrib>Yokoyama, Shin-ichiro</creatorcontrib><creatorcontrib>Yanase, Emiko</creatorcontrib><creatorcontrib>Niwa, Toshio</creatorcontrib><creatorcontrib>Ebihara, Akio</creatorcontrib><creatorcontrib>Inagaki, Mizuho</creatorcontrib><creatorcontrib>Yamaguchi, Keiichi</creatorcontrib><creatorcontrib>Kuwata, Kazuo</creatorcontrib><creatorcontrib>Kato, Yuta</creatorcontrib><creatorcontrib>Sakurada, Osamu</creatorcontrib><creatorcontrib>Suzuki, Tohru</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of bioscience and bioengineering</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kawada, Yuika</au><au>Goshima, Tomoko</au><au>Sawamura, Rie</au><au>Yokoyama, Shin-ichiro</au><au>Yanase, Emiko</au><au>Niwa, Toshio</au><au>Ebihara, Akio</au><au>Inagaki, Mizuho</au><au>Yamaguchi, Keiichi</au><au>Kuwata, Kazuo</au><au>Kato, Yuta</au><au>Sakurada, Osamu</au><au>Suzuki, Tohru</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Daidzein reductase of Eggerthella sp. YY7918, its octameric subunit structure containing FMN/FAD/4Fe-4S, and its enantioselective production of R-dihydroisoflavones</atitle><jtitle>Journal of bioscience and bioengineering</jtitle><addtitle>J Biosci Bioeng</addtitle><date>2018-09-01</date><risdate>2018</risdate><volume>126</volume><issue>3</issue><spage>301</spage><epage>309</epage><pages>301-309</pages><issn>1389-1723</issn><eissn>1347-4421</eissn><abstract>S-Equol is a metabolite of daidzein, a type of soy isoflavone, and three reductases are involved in the conversion of daidzein by specific intestinal bacteria. S-Equol is thought to prevent hormone-dependent diseases. We previously identified the equol producing gene cluster (eqlABC) of Eggerthella sp. YY7918. Daidzein reductase (DZNR), encoded by eqlA, catalyzes the reduction of daidzein to dihydrodaidzein (the first step of equol synthesis), which was confirmed using a recombinant enzyme produced in Escherichia coli. Here, we purified recombinant DZNR to homogeneity and analyzed its enzymological properties. DZNR contained FMN, FAD, and one 4Fe-4S cluster per 70-kDa subunit as enzymatic cofactors. DZNR reduced the CC bond between the C-2 and C-3 positions of daidzein, genistein, glycitein, and formononetin in the presence of NADPH. R-Dihydrodaidzein and R-dihydrogenistein were highly stereo-selectively produced from daidzein and genistein. The Km and kcat for daidzein were 11.9 μM and 6.7 s−1, and these values for genistein were 74.1 μM and 28.3 s−1, respectively. This enzyme showed similar kinetic parameters and wide substrate specificity for isoflavone molecules. Thus, this enzyme appears to be an isoflavone reductase. Gel filtration chromatography and chemical cross-linking analysis of the active form of DZNR suggested that the enzyme consists of an octameric subunit structure. We confirmed this by small-angle X-ray scattering and transmission electron microscopy at a magnification of ×200,000. DZNR formed a globular four-petal cloverleaf structure with a central vertical hole. The maximum particle size was 173 Å.</abstract><cop>Japan</cop><pub>Elsevier B.V</pub><pmid>29699942</pmid><doi>10.1016/j.jbiosc.2018.03.018</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-1763-9084</orcidid></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1389-1723 |
ispartof | Journal of bioscience and bioengineering, 2018-09, Vol.126 (3), p.301-309 |
issn | 1389-1723 1347-4421 |
language | eng |
recordid | cdi_proquest_miscellaneous_2032433027 |
source | Access via ScienceDirect (Elsevier) |
subjects | Equol FAD FMN Gut microflora Isoflavone NADPH Old yellow enzyme Reductase Small-angle X-ray scattering |
title | Daidzein reductase of Eggerthella sp. YY7918, its octameric subunit structure containing FMN/FAD/4Fe-4S, and its enantioselective production of R-dihydroisoflavones |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T05%3A46%3A01IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Daidzein%20reductase%20of%20Eggerthella%20sp.%20YY7918,%20its%20octameric%20subunit%20structure%20containing%20FMN/FAD/4Fe-4S,%20and%20its%20enantioselective%20production%20of%20R-dihydroisoflavones&rft.jtitle=Journal%20of%20bioscience%20and%20bioengineering&rft.au=Kawada,%20Yuika&rft.date=2018-09-01&rft.volume=126&rft.issue=3&rft.spage=301&rft.epage=309&rft.pages=301-309&rft.issn=1389-1723&rft.eissn=1347-4421&rft_id=info:doi/10.1016/j.jbiosc.2018.03.018&rft_dat=%3Cproquest_cross%3E2032433027%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2032433027&rft_id=info:pmid/29699942&rft_els_id=S1389172317309817&rfr_iscdi=true |