State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, People's Republic of China.
Appl Biochem Biotechnol. 2012 May;167(1):73-80. doi: 10.1007/s12010-012-9653-4. Epub 2012 Apr 14.
A novel biosensor strain (Escherichia coli ALM403) that responded to N-acyl homoserine lactone (AHL) was constructed using a luxR-Plux cassette as a regulatory sequence and β-mannanase as a reporter gene. Dinitrosalicylic acid method was used to detect the response of the sensor strain to N-acyl homoserine lactone. By investigating the response to a range of concentrations of N-β-oxooctanoyl-L-homoserine lactone (OOHL), it was demonstrated that the expression of mannanase in E. coli ALM403 could be greatly enhanced by OOHL and resulted in an assayable phenotype. A high-throughput screening approach was developed to isolate AHL-degrading microorganisms, and a marine Halomonas sp. S66-4 showing a marked AHL-degrading ability was successfully isolated. In conclusion, the bioassay system provided a simple and efficient approach to isolate AHL-degrading bacteria.
利用 luxR-Plux 盒作为调控序列，β-甘露聚糖酶作为报告基因，构建了一种能响应 N-酰基高丝氨酸内酯 (AHL) 的新型生物传感器菌株（大肠杆菌 ALM403）。采用二硝基水杨酸法检测传感器菌株对 N-酰基高丝氨酸内酯的响应。通过研究该传感器菌株对一系列 N-β-氧代辛酰基-L-高丝氨酸内酯 (OOHL) 浓度的响应，表明 OOHL 能极大地增强大肠杆菌 ALM403 中甘露聚糖酶的表达，从而产生可检测的表型。建立了一种高通量筛选方法来分离 AHL 降解微生物，成功分离到一株具有显著 AHL 降解能力的海洋盐单胞菌 S66-4。总之，该生物测定系统为分离 AHL 降解菌提供了一种简单有效的方法。
