Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase.
The quorum sensing (QS) systems in Gram-negative bacteria are mostly associated with diffusible N-acyl-L-homoserine lactones (AHLs). AHL-degrading enzymes hydrolyze the AHLs into inactive molecules, thereby blocking the QS systems that are closely linked to virulence factor production and biofilm formation. Consequently, these enzymes have recently attracted intense interest for the development of anti-infection therapies for plants and animals. However, despite significant progress in the investigation of AHL-degrading enzymes, no structure is yet available. Accordingly, this study reports on the expression and purification of the AHL-lactonase from Bacillus thuringiensis subsp. kurstaki HD263, as well as the successful crystallization of the enzyme. High-quality native crystals were obtained and a complete data set collected at 2.0 A resolution. The native crystal was found to belong to the space group P2(1)2(1)2(1), with unit cell parameters a=52.7 A, b=55.9 A, and c=74.1 A and one molecule in the asymmetric unit. MAD data were also collected at 2.4 A resolution for a SeMet-substituted crystal.
Kim
Myung Hee
MH
Korea Research Institute of Bioscience and Biotechnology, Yusong, Daejon 305-600, Republic of Korea.
Kang
Hye Ok
HO
Kang
Beom Sik
BS
Kim
Kyung-Jin
KJ
Choi
Won-Chan
WC
Oh
Tae-Kwang
TK
Lee
Choong Hwan
CH
Lee
Jung-Kee
JK
D001413:Bacillus thuringiensis / Q000201:enzymology*; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray
D002265:Carboxylic Ester Hydrolases; C432003:N-acyl homoserine lactonase