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

被引：9

作者：

Kumar, Pravindra ^{[1
,2
,3
]}

Gomez-Gil, Leticia ^{[4
]}

Mohammadi, Mahmood ^{[5
]}

Sylvestre, Michel ^{[5
]}

Eltis, Lindsay D. ^{[4
]}

Bolin, Jeffrey T. ^{[1
,2
]}

机构：

[1] Purdue Univ, Dept Biol Sci, W Lafayette, IN 47907 USA

[2] Purdue Univ, Ctr Canc Res, W Lafayette, IN 47907 USA

[3] Indian Inst Technol, Dept Biotechnol, Roorkee, Uttar Pradesh, India

[4] Univ British Columbia, Dept Microbiol, Inst Life Sci, Vancouver, BC V6T 1Z3, Canada

[5] Univ Quebec, Inst Armand Frappier, INRS, Laval, PQ H7V 1B7, Canada

来源：

ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2011年 / 67卷

基金：

加拿大自然科学与工程研究理事会; 美国国家卫生研究院;

关键词：

SP STRAIN LB400; POLYCHLORINATED-BIPHENYLS; DIOXYGENASE; DEGRADATION; PROTEINS; GROWTH; CHLOROBIPHENYLS; MUTAGENESIS; PATHWAY; ENZYMES;

D O I：

10.1107/S1744309110043393

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (similar to 8 angstrom) 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 angstrom resolution grew in space group P1. Crystals of the BPDOP4 and BPDORR41 variants of BPDOLB400 grew in space group P2(1).

引用

页码：59 / 62

页数：4

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