Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase

Hee Kim Myung; Ok Kang Hye; Sik Kang Beom; Kyung Jin Kim; Won Chan Choi; Tae Kwang Oh; Hwan Lee Choong; Jung Kee Lee

doi:10.1016/j.bbapap.2005.04.004

Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase

Hee Kim Myung, Ok Kang Hye, Sik Kang Beom, Kyung Jin Kim, Won Chan Choi, Tae Kwang Oh, Hwan Lee Choong, Jung Kee Lee

Korea Research Institute of Bioscience and Biotechnology

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

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 Å resolution. The native crystal was found to belong to the space group P2₁2₁2₁, with unit cell parameters a = 52.7 Å, b = 55.9 Å, and c = 74.1 Å and one molecule in the asymmetric unit. MAD data were also collected at 2.4 Å resolution for a SeMet-substituted crystal.

Original language	English
Pages (from-to)	5-8
Number of pages	4
Journal	Biochimica et Biophysica Acta - Proteins and Proteomics
Volume	1750
Issue number	1
DOIs	https://doi.org/10.1016/j.bbapap.2005.04.004
State	Published - 15 Jun 2005

Keywords

AHL-lactonase
Crystallization
N-acyl-L-homoserine lactone
Quorum sensing

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10.1016/j.bbapap.2005.04.004

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@article{d0135c1fdeac4df48b842e51ba0fc16e,

title = "Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase",

abstract = "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 {\AA} resolution. The native crystal was found to belong to the space group P212121, with unit cell parameters a = 52.7 {\AA}, b = 55.9 {\AA}, and c = 74.1 {\AA} and one molecule in the asymmetric unit. MAD data were also collected at 2.4 {\AA} resolution for a SeMet-substituted crystal.",

keywords = "AHL-lactonase, Crystallization, N-acyl-L-homoserine lactone, Quorum sensing",

author = "Myung, {Hee Kim} and Hye, {Ok Kang} and Beom, {Sik Kang} and Kim, {Kyung Jin} and Choi, {Won Chan} and Oh, {Tae Kwang} and Choong, {Hwan Lee} and Lee, {Jung Kee}",

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T1 - Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase

AU - Myung, Hee Kim

AU - Hye, Ok Kang

AU - Beom, Sik Kang

AU - Kim, Kyung Jin

AU - Choi, Won Chan

AU - Oh, Tae Kwang

AU - Choong, Hwan Lee

AU - Lee, Jung Kee

PY - 2005/6/15

Y1 - 2005/6/15

N2 - 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 Å resolution. The native crystal was found to belong to the space group P212121, with unit cell parameters a = 52.7 Å, b = 55.9 Å, and c = 74.1 Å and one molecule in the asymmetric unit. MAD data were also collected at 2.4 Å resolution for a SeMet-substituted crystal.

AB - 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 Å resolution. The native crystal was found to belong to the space group P212121, with unit cell parameters a = 52.7 Å, b = 55.9 Å, and c = 74.1 Å and one molecule in the asymmetric unit. MAD data were also collected at 2.4 Å resolution for a SeMet-substituted crystal.

KW - AHL-lactonase

KW - Crystallization

KW - N-acyl-L-homoserine lactone

KW - Quorum sensing

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Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase

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