文化大學機構典藏 CCUR:Item 987654321/29214
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    题名: Bioconversion of Pinoresinol into Matairesinol by Use of Recombinant Escherichia coli
    作者: Kuo, Han-Jung
    Wei, Zhi-Yu
    Lu, Pei-Chun
    Huang, Pung-Ling
    Lee, Kung-Ta
    贡献者: 生物科技研究所
    关键词: BENZYLIC ETHER REDUCTASES
    ISOFLAVONE REDUCTASE
    SECOISOLARICIRESINOL DEHYDROGENASE
    LARICIRESINOL REDUCTASES
    BIFUNCTIONAL ENZYME
    CRYSTAL-STRUCTURES
    MOLECULAR-CLONING
    FUSION PROTEIN
    LIGNANS
    PHYTOESTROGENS
    日期: 2014-05
    上传时间: 2015-01-26 15:51:01 (UTC+8)
    摘要: Lignans, a class of dimeric phenylpropanoid derivative found in plants, such as whole grains and sesame and flax seeds, have anticancer activity and can act as phytoestrogens. The lignans secoisolariciresinol and matairesinol can be converted in the mammalian proximal colon into enterolactone and enterodiol, respectively, which reduce the risk of breast and colon cancer. To establish an efficient bioconversion system to generate matairesinol from pinoresinol, the genes encoding pinoresinol-lariciresinol reductase (PLR) and secoisolariciresinol dehydrogenase (SDH) were cloned from Podophyllum pleianthum Hance, an endangered herb in Taiwan, and the recombinant proteins, rPLR and rSDH, were expressed in Escherichia coli and purified. The two genes, termed plr-PpH and sdh-PpH, were also linked to form two bifunctional fusion genes, plr-sdh and sdh-plr, which were also expressed in E. coli and purified. Bioconversion in vitro at 22 C for 60 min showed that the conversion efficiency of fusion protein PLR-SDH was higher than that of the mixture of rPLR and rSDH. The percent conversion of (+)-pinoresinol to matairesinol was 49.8% using PLR-SDH and only 17.7% using a mixture of rPLR and rSDH. However, conversion of (+)-pinoresinol by fusion protein SDH-PLR stopped at the intermediate product, secoisolariciresinol. In vivo, (+)-pinoresinol was completely converted to matairesinol by living recombinant E. coli expressing PLR-SDH without addition of cofactors.
    關聯: APPLIED AND ENVIRONMENTAL MICROBIOLOGY 卷: 80 期: 9 頁碼: 2687-2692
    显示于类别:[生物科技研究所 ] 期刊論文

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