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j.bbrc.2010.12.028.pdf | 110.19 kB | Adobe PDF | 見る/開く |
タイトル: | A novel strictly NADPH-dependent Pichia stipitis xylose reductase constructed by site-directed mutagenesis. |
著者: | Khattab, Sadat Mohammad Rezq Watanabe, Seiya Saimura, Masayuki Kodaki, Tsutomu |
著者名の別形: | 小瀧, 努 |
キーワード: | Coenzyme specificity Xylose reductase Site-directed mutagenesis |
発行日: | 14-Jan-2011 |
出版者: | Elsevier Inc. |
誌名: | Biochemical and biophysical research communications |
巻: | 404 |
号: | 2 |
開始ページ: | 634 |
終了ページ: | 637 |
抄録: | Xylose reductase (XR) and xylitol dehydrogenase (XDH) are the key enzymes for xylose fermentation and have been widely used for construction of a recombinant xylose fermenting yeast. The effective recycling of cofactors between XR and XDH has been thought to be important to achieve effective xylose fermentation. Efforts to alter the coenzyme specificity of XR and HDX by site-directed mutagenesis have been widely made for improvement of efficiency of xylose fermentation. We previously succeeded by protein engineering to improve ethanol production by reversing XDH dependency from NAD(+) to NADP(+). In this study, we applied protein engineering to construct a novel strictly NADPH-dependent XR from Pichia stipitis by site-directed mutagenesis, in order to recycle NADPH between XR and XDH effectively. One double mutant, E223A/S271A showing strict NADPH dependency with 106% activity of wild-type was generated. A second double mutant, E223D/S271A, showed a 1.27-fold increased activity compared to the wild-type XR with NADPH and almost negligible activity with NADH. |
著作権等: | © 2010 Elsevier Inc. This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 |
URI: | http://hdl.handle.net/2433/134797 |
DOI(出版社版): | 10.1016/j.bbrc.2010.12.028 |
PubMed ID: | 21146502 |
出現コレクション: | 学術雑誌掲載論文等 |
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