WEKO3
-
RootNode
アイテム
Camphor pathway redux: functional recombinant expression of 2,5- and 3,6-diketocamphane monooxygenases of Pseudomonas putida ATCC 17453 with their cognate flavin reductase catalyzing Baeyer-Villiger reactions
http://hdl.handle.net/10112/8080
http://hdl.handle.net/10112/8080cf7e8297-ed4a-4aee-9275-9b4d8ba4f296
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
KU-1100-20130300-11.pdf (5.7 MB)
|
|
| Item type | 学術雑誌論文 / Journal Article(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2014-01-10 | |||||
| タイトル | ||||||
| タイトル | Camphor pathway redux: functional recombinant expression of 2,5- and 3,6-diketocamphane monooxygenases of Pseudomonas putida ATCC 17453 with their cognate flavin reductase catalyzing Baeyer-Villiger reactions | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
| 資源タイプ | journal article | |||||
| 著者 |
岩木, 宏明
× 岩木, 宏明× Grosse, Stephan× Bergeron, Helene× Leisch, Hannes× Morley, Krista× 長谷川, 喜衛× Lau, Peter C.K |
|||||
| 著者別名 | ||||||
| 識別子Scheme | WEKO | |||||
| 識別子 | 1854 | |||||
| 姓名 | 岩木, 宏明 | |||||
| 概要 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | Whereas the biochemical properties of the monooxygenase components that catalyze the oxidation of 2,5-diketocamphane and 3,6-diketocamphane (2,5-DKCMO and 3,6-DKCMO, respectively) in the initial catabolic steps of (+) and (−) isomeric forms of camphor metabolism in Pseudomonas putida ATCC 17453 are relatively well characterized, the actual identity of the flavin reductase (Fred) component that provides the reduced flavin to the oxygenases is hitherto ill-defined. In this study, a 37-kDa Fred was purified from camphor-induced culture of P. putida ATCC 17453 and this facilitated cloning and characterization of the requisite protein. The active Fred is a homodimer with a subunit molecular mass of 18-kDa that uses NADH as electron donor (Km = 32 μM) and it catalyzes the reduction of FMN (Km = 3.6 μM; kcat = 283 s-1) in preference to FAD (Km = 19 μM; kcat = 128 s-1). Sequence determination of ∼40-kb of the camphor (CAM) degradation plasmid revealed the locations of two isofunctional 2,5-DKCMO genes (camE25-1 for 2,5-DKCMO-1, and camE25-2 for 2,5-DKCMO-2) as well as that of 3,6-DKCMO-encoding gene (camE36). In addition, by pulsed-field gel electrophoresis, the CAM plasmid was established to be linear and ∼533-kb in length. To enable functional assessment of the two-component monooxygenase system in Baeyer-Villiger oxidations, recombinant plasmids expressing Fred in tandem with the respective 2,5-DKCMO and 3,6-DKCMO encoding genes in Escherichia coli were constructed. Comparative substrate profiling of the isofunctional 2,5-DCKMOs did not yield obvious differences in Baeyer-Villiger biooxidations but they are distinct from 3,6-DKCMO in the stereoselective oxygenations with various mono- and bicyclic ketone substrates. | |||||
| 書誌情報 |
Applied and Environmental Microbiology 発行日 2013-03 |
|||||
| ISSN | ||||||
| 収録物識別子タイプ | ISSN | |||||
| 収録物識別子 | 00992240 | |||||
| DOI | ||||||
| 関連タイプ | isVersionOf | |||||
| 識別子タイプ | DOI | |||||
| 関連識別子 | 10.1128/AEM.03958-12 | |||||
| 権利 | ||||||
| 権利情報 | (C)American Society for Microbiology: This ariticle is available at http://aem.asm.org/content/early/2013/03/18/AEM.03958-12 | |||||
| 著者版フラグ | ||||||
| 出版タイプ | AM | |||||
| 出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
| 出版者 | ||||||
| 出版者 | American Society for Microbiology | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | 平成24年度関西大学若手研究者育成経費 | |||||
Share
Cite as
岩木, 宏明, Grosse, Stephan, Bergeron, Helene, Leisch, Hannes, Morley, Krista, 長谷川, 喜衛, Lau, Peter C.K, 2013, Camphor pathway redux: functional recombinant expression of 2,5- and 3,6-diketocamphane monooxygenases of Pseudomonas putida ATCC 17453 with their cognate flavin reductase catalyzing Baeyer-Villiger reactions: American Society for Microbiology.
Loading...
