Anaerobic crystallization and initial X-ray diffraction data of biphenyl 2,3-dioxygenase from Burkholderia xenovorans LB400: addition of agarose improved the quality of the crystals.

Pravindra Kumar (1, 2) , Leticia Gómez-Gil (3) , Mahmood Mohammadi (4) , Michel Sylvestre (4) , Lindsay D Eltis (3) , Jeffrey T Bolin (1)

1 Department of Biological Sciences [West Lafayette]
2 Department of Biotechnology
3 Department of Microbiology
4 INRS-IAF - Institut Armand Frappier

Biphenyl 2,3-dioxygenase (BPDO; EC 1.14.12.18) catalyzes the initial step in the degradation of biphenyl and some polychlorinated biphenyls (PCBs). BPDOLB400, the terminal dioxygenase component from Burkholderia xenovorans LB400, a proteobacterial species that degrades a broad range of PCBs, has been crystallized under anaerobic conditions by sitting-drop vapour diffusion. Initial crystals obtained using various polyethylene glycols as precipitating agents diffracted to very low resolution (∼8 Å) and the recorded reflections were diffuse and poorly shaped. The quality of the crystals was significantly improved by the addition of 0.2% agarose to the crystallization cocktail. In the presence of agarose, wild-type BPDOLB400 crystals that diffracted to 2.4 Å resolution grew in space group P1. Crystals of the BPDOP4 and BPDORR41 variants of BPDOLB400 grew in space group P2(1).

Domaines: Biotechnologies; Biotechnologie

Pravindra Kumar, Leticia Gómez-Gil, Mahmood Mohammadi, Michel Sylvestre, Lindsay D Eltis, et al.. Anaerobic crystallization and initial X-ray diffraction data of biphenyl 2,3-dioxygenase from Burkholderia xenovorans LB400: addition of agarose improved the quality of the crystals.. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 2011, 67 (Pt 1), pp.59-62. ⟨10.1107/S1744309110043393⟩. ⟨pasteur-00723272⟩