BioDesign 2022; 10(1): 12-15
Published online March 30, 2022
© Korean Society for Structural Biology
Jihye Seok1 and Kyung-Jin Kim1,2,*
1School of Life Sciences, BK21 FOUR KNU Creative BioResearch Group, Kyungpook National University, Daegu 41566, Korea
2KNU Institute for Microorganisms, Kyungpook National University, Daegu 41566, Korea
Correspondence to: *firstname.lastname@example.org
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 = 0°, γ = 120°. With six molecules of PpFerB per asymmetric unit, the crystal volume per unit of protein mass is 2.37 Å3 Da–1, corresponding to a solvent content of approximately 48.08%.