Department of Material and Environmental Chemistry, Graduate School of Engineering, Utsunomiya University, 7-1-2 Yoto, Utsunomiya 321-8585, Japan.
J Biosci Bioeng. 2012 Jan;113(1):20-5. doi: 10.1016/j.jbiosc.2011.09.006. Epub 2011 Oct 22.
N-Acylhomoserine lactones (AHLs) are used as quorum-sensing signal molecules by many gram-negative bacteria. We have reported that Solibacillus silvestris, which was isolated from the potato leaf, has AHL-degrading activity. To identify the AHL-degrading gene, whole genome sequencing of S. silvestris StLB046 was performed by using pyrosequencing technology. As the result of the BLAST search, one predicted ORF (ahlS) showed slight similarity to AiiA-like AHL lactonase from Bacillus cereus group. Escherichia coli harboring the ahlS-expressing plasmid showed high AHL-degrading activity. The ahlS-cording region was also amplified by PCR from the other potato leaf-associated and AHL-degrading S. silvestris strains. Purified AhlS as a maltose binding fusion protein showed high AHL-degrading activity and catalyzes AHL ring opening by hydrolyzing lactones. In addition, expression of ahlS in plant pathogen Pectobacterium carotovorum subsp. carotovorum attenuated maceration of the potato slices. Our results suggest that AHL-degrading activity of ahlS might perform useful functions such as useful biocontrol agents.
N-酰基高丝氨酸内酯 (AHLs) 被许多革兰氏阴性菌用作群体感应信号分子。我们曾报道,从马铃薯叶片中分离出的 Solibacillus silvestris 具有 AHL 降解活性。为了鉴定 AHL 降解基因,我们使用焦磷酸测序技术对 S. silvestris StLB046 进行了全基因组测序。通过 BLAST 搜索,一个预测的 ORF (ahlS) 显示与芽孢杆菌属 AiiA 样 AHL 内酯酶有轻微相似性。携带 ahlS 表达质粒的大肠杆菌表现出高 AHL 降解活性。从其他与马铃薯叶片相关且具有 AHL 降解活性的 S. silvestris 菌株中也通过 PCR 扩增了 ahlS 编码区。作为麦芽糖结合融合蛋白纯化的 AhlS 表现出高 AHL 降解活性,并通过水解内酯催化 AHL 开环。此外,在植物病原菌 Pectobacterium carotovorum subsp. carotovorum 中表达 ahlS 可减轻马铃薯切片的软化。我们的结果表明,ahlS 的 AHL 降解活性可能具有有用的功能,例如作为有用的生物防治剂。