Reference - Detail
| RRC ID | 5705 |
|---|---|
| Author | Sato S, Nomura CT, Abe H, Doi Y, Tsuge T. |
| Title | Poly[(R)-3-hydroxybutyrate] formation in Escherichia coli from glucose through an enoyl-CoA hydratase-mediated pathway. |
| Journal | 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. |
| Volume | 103(1) |
| Pages | 38-44 |
| Published | 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 |
| Times Cited | 19 |
| WOS Category | FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 143 |
| Altmetric score changes over past 6months | 3.0 |
| Resource | |
| Prokaryotes E. coli | CAG18497 |