Department of Material and Environmental Chemistry, Graduate School of Engineering, Utsunomiya University, 7-1-2 Yoto, Utsunomiya 321-8585, Japan.
Appl Environ Microbiol. 2010 Apr;76(8):2524-30. doi: 10.1128/AEM.02738-09. Epub 2010 Feb 19.
N-Acylhomoserine lactones (AHLs) are used as quorum-sensing signal molecules by many Gram-negative bacteria. We have reported that Microbacterium testaceum StLB037, which was isolated from the leaf surface of potato, has AHL-degrading activity. In this study, we cloned the aiiM gene from the genomic library of StLB037, which has AHL-degrading activity and shows high homology with the alpha/beta hydrolase fold family from Actinobacteria. Purified AiiM as a maltose binding fusion protein showed high degrading activity of AHLs with both short- and long-chain AHLs with or without substitution at carbon 3. High-performance liquid chromatography analysis revealed that AiiM works as an AHL lactonase that catalyzes AHL ring opening by hydrolyzing lactones. In addition, expression of AiiM in the plant pathogen Pectobacterium carotovorum subsp. carotovorum reduced pectinase activity markedly and attenuated soft rot symptoms on potato slices. In conclusion, this study indicated that AiiM might be effective in quenching quorum sensing of P. carotovorum subsp. carotovorum.
N-酰基高丝氨酸内酯 (AHLs) 被许多革兰氏阴性细菌用作群体感应信号分子。我们曾报道过,从马铃薯叶表面分离出的微杆菌 StLB037 具有 AHL 降解活性。在本研究中,我们从 StLB037 的基因组文库中克隆了aiiM 基因,该基因具有 AHL 降解活性,与放线菌的 α/β 水解酶折叠家族具有高度同源性。作为麦芽糖结合融合蛋白纯化的 AiiM 对具有或不具有碳 3 取代的短链和长链 AHL 均表现出高降解活性。高效液相色谱分析表明,AiiM 作为 AHL 内酯酶起作用,通过水解内酯催化 AHL 环的打开。此外,在植物病原菌果胶杆菌亚种胡萝卜软腐病菌中表达 AiiM 可显著降低果胶酶活性并减轻马铃薯切片上的软腐病症状。总之,本研究表明 AiiM 可能有效抑制果胶杆菌亚种胡萝卜软腐病菌的群体感应。