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

Yoshikazu Shimada; Setsuko Yasuda; Masayuki Takahashi; Takashi Hayashi; Norihiro Miyazawa; Ikutaro Sato

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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

机译：新型NADP（H）依赖性黄豆苷元还原酶的克隆和表达，该酶涉及黄豆产生素的代谢酶，其来自产生赤霉素的乳球菌菌株20-92。

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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 purified a novel NADP(H)-dependent daidzein reductase (L-DZNR) from Lactococcus strain 20-92 (Lactococcus 20-92; S. Uchiyama, T. Ueno, and T. Suzuki, international patent WO2005/000042) that is involved in the metabolism of soy isoflavones and equol production and converts daidzein to dihydrodaidzein. Partial amino acid sequences were determined from purified L-DZNR, and the gene encoding L-DZNR was cloned. The nucleotide sequence of this gene consists of an open reading frame of 1,935 nucleotides, and the deduced amino acid sequence consists of 644 amino acids. L-DZNR contains two cofactor binding motifs and an 4Fe-4S cluster. 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.

机译：紫杉醇是黄豆苷元通过肠道菌群产生的代谢产物，由于它对几种性激素依赖性疾病（例如，更年期疾病和较低的骨密度）的保护或改善能力而倍受青睐，因此引起了极大的关注。其他异黄酮。我们从乳球菌 20-92（乳球菌 20-92; S.Uchiyama，T.Ueno）纯化了一种新型的NADP（H）依赖性黄豆苷元还原酶（L-DZNR）。和T. Suzuki，国际专利WO2005 / 000042），其涉及大豆异黄酮的代谢和雌马酚的产生，并将大豆苷元转化为二氢黄豆苷元。从纯化的L-DZNR确定部分氨基酸序列，并克隆编码L-DZNR的基因。该基因的核苷酸序列由1,935个核苷酸的开放阅读框组成，推导的氨基酸序列由644个氨基酸组成。 L-DZNR包含两个辅因子结合基序和一个4Fe-4S簇。进一步认为，L-DZNR是属于 o ld y ellow e nzyme（OYE）家庭。重组组氨酸标签的L-DZNR在大肠杆菌中表达。重组蛋白将黄豆苷元转化为对映体选择性的（ S ）-二氢黄豆苷元。这是首次分离与大豆苷元代谢和肠细菌中雌马酚产生有关的酶的报告。 展开▼

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《Applied and Environmental Microbiology》 |2010年第17期|共10页

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Yoshikazu Shimada; Setsuko Yasuda; Masayuki Takahashi; Takashi Hayashi; Norihiro Miyazawa; Ikutaro Sato;
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1. 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 [J] . Yoshikazu Shimada, Setsuko Yasuda, Masayuki Takahashi, Applied and Environmental Microbiology . 2010,第17期

机译：新型NADP（H）依赖性黄豆苷元还原酶的克隆和表达，该酶涉及黄豆产生素的代谢酶，其来自产生赤霉素的乳球菌菌株20-92。

2. 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 [J] . Yoshikazu Shimada, Setsuko Yasuda, Masayuki Takahashi, Applied Microbiology . 2010,第17期

机译：新型NADP（H）依赖性黄豆苷元还原酶的克隆和表达，该酶涉及黄豆产生素的代谢酶，其来自产生赤霉素的乳球菌菌株20-92。

3. 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 [J] . Yoshikazu Shimada, Setsuko Yasuda, Masayuki Takahashi, Applied Microbiology . 2010,第17期

机译：新型NADP（H）依赖性黄豆苷元还原酶的克隆和表达，该酶涉及黄豆产生素的代谢酶，其来自产生赤霉素的乳球菌菌株20-92。

4. SPECIFICITY FOR THE COENZYME IN FERREDOXIN-NADP~+ REDUCTASE: AN ATTEMPT TO CREATE A NAD~+/H DEPENDENT ENZYME [C] . Marta Martinez-Julvez, Jesus Tejero, Jose Ramon Peregrina, International Congress of Photosynthesis . 2005

机译：Ferriedoxin-NADP〜+还原酶中辅酶的特异性：试图产生NAD〜+ / H依赖性酶

5. Structural studies of three NAD(P)H-dependent enzymes involved in sugar metabolism: Mannitol dehydrogenase, xylose reductase, and lactate dehydrogenase. [D] . Kavanagh, Kathryn Louise. 2003

机译：糖代谢中涉及的三种NAD（P）H依赖酶的结构研究：甘露醇脱氢酶，木糖还原酶和乳酸脱氢酶。

6. 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 [O] . Yoshikazu Shimada, Setsuko Yasuda, Masayuki Takahashi, 2010

机译：新型NADP（H）依赖性黄豆苷元还原酶的克隆和表达该酶涉及黄豆产生素的代谢酶其来自产生赤霉素的乳球菌菌株20-92。

7. 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▿ [O] . Shimada, Yoshikazu, Yasuda, Setsuko, Takahashi, Masayuki, 2010

机译：从产生赤霉素的乳球菌菌株20-92▿克隆和表达一种新型的依赖NADP（H）的大豆苷元还原酶，一种涉及大豆苷元代谢的酶。

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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

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