title: 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.
abstract: 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).
authors.affiliation: Department of Biological Sciences and Center for Cancer Research, Purdue University, West Lafayette, Indiana, USA. kumarfbs@iitr.ernet.in
authors.forename: Pravindra
authors.initials: P
authors.lastname: Kumar
authors.forename: Leticia
authors.initials: L
authors.lastname: Gómez-Gil
authors.forename: Mahmood
authors.initials: M
authors.lastname: Mohammadi
authors.forename: Michel
authors.initials: M
authors.lastname: Sylvestre
authors.forename: Lindsay D
authors.initials: LD
authors.lastname: Eltis
authors.forename: Jeffrey T
authors.initials: JT
authors.lastname: Bolin
mesh_terms: D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000737:chemistry*; D019117:Burkholderia / Q000201:enzymology*; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D012685:Sepharose; D010105:Oxygenases; C074161:biphenyl-2,3-dioxygenase