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タイトル: | Crucial role of 4-deoxy-L-erythro-5-hexoseulose uronate reductase for alginate utilization revealed by adaptive evolution in engineered Saccharomyces cerevisiae |
著者: | Matsuoka, Fumiya Hirayama, Makoto Kashihara, Takayuki Tanaka, Hideki Hashimoto, Wataru https://orcid.org/0000-0003-0185-2371 (unconfirmed) Murata, Kousaku Kawai, Shigeyuki |
著者名の別形: | 松岡, 史也 橋本, 渉 村田, 幸作 河井, 重幸 |
発行日: | 23-Jun-2017 |
出版者: | Springer Nature |
誌名: | Scientific Reports |
巻: | 7 |
論文番号: | 4206 |
抄録: | In brown macroalgae, alginate and D-mannitol are promising carbohydrates for biorefinery. Saccharomyces cerevisiae is widely used as a microbial cell factory, but this budding yeast is unable to utilize either alginate or D-mannitol. Alginate can be depolymerized by both endo-type and exo-type alginate lyases, yielding a monouronate, 4-deoxy-L-erythro-5-hexoseulose uronate (DEH), a key intermediate in the metabolism of alginate. Here, we constructed engineered two S. cerevisiae strains that are able to utilize both DEH and D-mannitol on two different strain backgrounds, and we also improved their aerobic growth in a DEH liquid medium through adaptive evolution. In both evolved strains, one of the causal mutations was surprisingly identical, a c.50A > G mutation in the codon-optimized NAD(P)H-dependent DEH reductase gene, one of the 4 genes introduced to confer the capacity to utilize DEH. This mutation resulted in an E17G substitution at a loop structure near the coenzyme-binding site of this reductase, and enhanced the reductase activity and aerobic growth in both evolved strains. Thus, the crucial role for this reductase reaction in the metabolism of DEH in the engineered S. cerevisiae is demonstrated, and this finding provides significant information for synthetic construction of a S. cerevisiae strain as a platform for alginate utilization. |
記述: | 代謝改変酵母のアルギン酸モノマー代謝能向上メカニズムの解明--国産海洋バイオマス資源から代替ガソリンや合成ゴム原料を生産するための重要な第一歩--. 京都大学プレスリリース. 2017-06-26. |
著作権等: | © The Author(s) 2017. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
URI: | http://hdl.handle.net/2433/226165 |
DOI(出版社版): | 10.1038/s41598-017-04481-3 |
PubMed ID: | 28646149 |
関連リンク: | https://www.kyoto-u.ac.jp/ja/research-news/2017-06-26-0 |
出現コレクション: | 学術雑誌掲載論文等 |
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