論文 - 詳細
| RRC ID | 5705 |
|---|---|
| 著者 | Sato S, Nomura CT, Abe H, Doi Y, Tsuge T. |
| タイトル | Poly[(R)-3-hydroxybutyrate] formation in Escherichia coli from glucose through an enoyl-CoA hydratase-mediated pathway. |
| ジャーナル | J Biosci Bioeng |
| Abstract |
In this study, a new metabolic pathway for the synthesis of poly[(R)-3-hydroxybutyrate] [P(3HB)] was constructed in a recombinant Escherichia coli strain that utilized forward and reverse reactions catalyzed by two substrate-specific enoyl-CoA hydratases, R-hydratase (PhaJ) and S-hydratase (FadB), to epimerize (S)-3HB-CoA to (R)-3HB-CoA via a crotonyl-CoA intermediate. The R-hydratase gene (phaJ(Ac)) from Aeromonas caviae was coexpressed with the PHA synthase gene (phaC(Re)) and 3-ketothiolase gene (phaA(Re)) from Ralstonia eutropha in fadR mutant E. coli strains (CAG18497 and LS5218), which had constitutive levels of the beta-oxidation multienzyme FadB(Ec). When grown on glucose as the sole carbon source, the cells accumulated P(3HB) up to an amount 6.5 wt% of the dry cell weight, whereas the control cells without phaJ(Ac) or fadR mutation accumulated significantly smaller amounts of P(3HB). These results suggest that PhaJ(Ac) and FadB(Ec) played an important role in supplying monomers for P(3HB) synthesis in the pathway. Furthermore, by using this pathway, a P(3HB)-concentration-dependent fluorescent staining screening technique was developed to rapidly identify cells that possess active R-hydratase. |
| 巻・号 | 103(1) |
| ページ | 38-44 |
| 公開日 | 2007-1-1 |
| DOI | 10.1263/jbb.103.38 |
| PII | S1389-1723(07)70022-1 |
| PMID | 17298899 |
| MeSH | Aeromonas / enzymology* Aeromonas / genetics Cupriavidus necator / enzymology* Cupriavidus necator / genetics Enoyl-CoA Hydratase / genetics Enoyl-CoA Hydratase / metabolism* Escherichia coli / genetics Escherichia coli / metabolism* Genetic Enhancement Glucose / metabolism* Hydroxybutyrates / metabolism* Polyesters / metabolism* Protein Engineering / methods* Recombinant Proteins / metabolism Signal Transduction / physiology |
| IF | 2.366 |
| 引用数 | 19 |
| WOS 分野 | FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 143 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 3.0 |
| リソース情報 | |
| 原核生物(大腸菌) | CAG18497 |