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KCI등재 학술저널

Purification, crystallization and X-ray crystallographic analysis of feruloyl-CoA hydratase/lyase of Pseudomonas putida KT2440

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Feruloyl-CoA hydratase/lyase (FerB) is an enzyme that catalyzes the final step of converting ferulic acid to biovanillin. In this study, we overexpressed FerB from Pseudomonas putida KT2440 (PpFerB) and purified the protein by Ni-NTA affinity and size-exclusion chromatography. The PpFerB protein was crystallized using hanging-drop vapor-diffusion method in the presence of 25% 1,2-propanediol, 0.1 M sodium phosphate dibasic/citric acid pH 4.2, 5% (w/v) PEG 3000, and 10% (v/v) glycerol at 293 K. X-ray diffraction data were collected to a maximum resolution of 1.65 Å. The PpFerB crystals belong to the space group R3 with unit cell parameters a = 87.7 Å, b = 87.7 Å, c = 198.0 Å, α = 90°, β = 90°, γ = 120°. With six molecules of PpFerB per asymmetric unit, the crystal volume per unit of protein mass is 2.37 ų Da-1, corresponding to a solvent content of approximately 48.08%.

INTRODUCTION

MATERIALS AND METHODS

RESULTS AND DISCUSSION

REFERENCES

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