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
リソース情報
原核生物(大腸菌) CAG18497