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H16_B0756 from Cupriavidus necator H16 is a putative enoyl-coenzyme A (CoA) hydratase/isomerase-family protein.
Enoyl-CoA hydratase (ECH) in β-oxidation might provide hydroxyalkanoate monomers for PHA production. However,β-oxidation pathway and ECH family enzymes in C. necator have not been fully characterized. To identify the function ofH16_B0756, the protein was overexpressed and purified to homogeneity by affinity and size-exclusion chromatography.
The H16_B0756 protein was crystallized using hanging-drop vapor-diffusion method in the presence of 30% polyethyleneglycol 550 monomethyl ether, 0.1 M sodium chloride and 0.1 M bicine, pH 9.0 at 295 K. X-ray diffraction data were collectedto a maximum resolution of 2.0 A. The crystal belonged to space group P3, with unit cell parameters a = b = 132.94 A, c =44.16, α = β = 90.0°, γ = 120.0°. With two molecules per asymmetric unit, the crystal volume per unit protein mass was 1.94A3 Da-1, which correspond to a solvent content of approximately 36.80%.
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참고문헌 (15)
참고문헌 신청
Benning, M. M / 2000 / New reactions in the crotonase superfamily : Structure of methylmalonyl CoA decarboxylase from Escherichia coli / Biochemistry 39:4630~4639
Budde, C, F / 2011 / Production of Poly(3-Hydroxybutyrate-co-3-Hydroxyhexanoate)from Plant Oil by Engineered Ralstonia eutropha Strains / Appl Environ Microbio 77:2847~2854
Chen, J. S / 2015 / Production of fatty acids in Ralstonia eutropha H16 by engineering beta-oxidation and carbon storage / PeerJ 3:e1468
Emsley, P / 2004 / Coot : model-building tools for molecular graphics / Acta Crystallogr D Biol Crystallogr 60:2126~2132
Fukui, T / 1998 / Expression and characterization of(R)-specific enoyl coenzyme A hydratase involved in polyhydroxyalkanoate biosynthesis by Aeromonas caviae / J Bacteriol 180:667~673
Budde, C, F / 2011 / Production of Poly(3-Hydroxybutyrate-co-3-Hydroxyhexanoate)from Plant Oil by Engineered Ralstonia eutropha Strains / Appl Environ Microbio 77:2847~2854
Chen, J. S / 2015 / Production of fatty acids in Ralstonia eutropha H16 by engineering beta-oxidation and carbon storage / PeerJ 3:e1468
Emsley, P / 2004 / Coot : model-building tools for molecular graphics / Acta Crystallogr D Biol Crystallogr 60:2126~2132
Fukui, T / 1998 / Expression and characterization of(R)-specific enoyl coenzyme A hydratase involved in polyhydroxyalkanoate biosynthesis by Aeromonas caviae / J Bacteriol 180:667~673
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