WEKO3
アイテム
Protein Engineering of the Transcriptional Activator FhlA To Enhance Hydrogen Production in Escherichia coli
http://hdl.handle.net/10228/00006289
http://hdl.handle.net/10228/000062899f25f7a5-42a5-4f75-8b36-e54f4ebdc599
| 名前 / ファイル | ライセンス | アクション |
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5639.full.pdf (580.2 kB)
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| アイテムタイプ | 学術雑誌論文 = Journal Article(1) | |||||||||||||
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| 公開日 | 2017-08-09 | |||||||||||||
| 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 | |||||||||||||
| タイトル | ||||||||||||||
| タイトル | Protein Engineering of the Transcriptional Activator FhlA To Enhance Hydrogen Production in Escherichia coli | |||||||||||||
| 言語 | ||||||||||||||
| 言語 | eng | |||||||||||||
| 著者 |
Sanchez-Torres, Viviana
× Sanchez-Torres, Viviana× 前田, 憲成
WEKO
649
× Wood, Thomas K. |
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| 抄録 | ||||||||||||||
| 内容記述タイプ | Abstract | |||||||||||||
| 内容記述 | Escherichia coli produces H2 from formate via the formate hydrogenlyase (FHL) complex during mixed acid fermentation; the FHL complex consists of formate dehydrogenase H (encoded by fdhF) for forming 2H+, 2e−, and CO2 from formate and hydrogenase 3 (encoded by hycGE) for synthesizing H2 from 2H+ and 2e−. FHL protein production is activated by the σ54 transcriptional activator FhlA, which activates transcription of fdhF and the hyc, hyp, and hydN-hypF operons. Here, through random mutagenesis using error-prone PCR over the whole gene, as well as over the fhlA region encoding the first 388 amino acids of the 692-amino-acid protein, we evolved FhlA to increase H2 production. The amino acid replacements in FhlA133 (Q11H, L14V, Y177F, K245R, M288K, and I342F) increased hydrogen production ninefold, and the replacements in FhlA1157 (M6T, S35T, L113P, S146C, and E363K) increased hydrogen production fourfold. Saturation mutagenesis at the codons corresponding to the amino acid replacements in FhlA133 and at position E363 identified the importance of position L14 and of E363 for the increased activity; FhlA with replacements L14G and E363G increased hydrogen production (fourfold and sixfold, respectively) compared to FhlA. Whole-transcriptome and promoter reporter constructs revealed that the mechanism by which the FhlA133 changes increase hydrogen production is by increasing transcription of all of the genes activated by FhlA (the FHL complex). With FhlA133, transcription of PfdhF and Phyc is less sensitive to formate regulation, and with FhlA363 (E363G), Phyc transcription increases but Phyp transcription decreases and hydrogen production is less affected by the repressor HycA. | |||||||||||||
| 書誌情報 |
Applied and Environmental Microbiology 巻 75, 号 17, p. 5639-5646, 発行日 2009-09 |
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| 出版者 | American Society for Microbiology | |||||||||||||
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| 関連タイプ | isIdenticalTo | |||||||||||||
| 識別子タイプ | DOI | |||||||||||||
| 関連識別子 | info:doi/10.1128/AEM.00638-09 | |||||||||||||
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| 収録物識別子タイプ | ISSN | |||||||||||||
| 収録物識別子 | 0099-2240 | |||||||||||||
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| 収録物識別子タイプ | ISSN | |||||||||||||
| 収録物識別子 | 1098-5336 | |||||||||||||
| 著作権関連情報 | ||||||||||||||
| 権利情報 | American Society for Microbiology | |||||||||||||
| 出版タイプ | ||||||||||||||
| 出版タイプ | VoR | |||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||||
| 査読の有無 | ||||||||||||||
| 値 | yes | |||||||||||||
| 連携ID | ||||||||||||||
| 値 | 880 | |||||||||||||
| 情報源 | ||||||||||||||
| 識別子タイプ | URI | |||||||||||||
| 関連識別子 | http://aem.asm.org/cgi/content/abstract/75/17/5639 | |||||||||||||
