title: A novel bioassay for high-throughput screening microorganisms with N-acyl homoserine lactone degrading activity.
abstract: 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.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, People's Republic of China.
authors.forename: Pengfu
authors.initials: P
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Gao
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Zongze
authors.initials: Z
authors.lastname: Shao
authors.forename: Jiping
authors.initials: J
authors.lastname: Shi
authors.forename: Ziduo
authors.initials: Z
authors.lastname: Liu
mesh_terms: D054742:Acyl-Butyrolactones / Q000378:metabolism*; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D044902:beta-Mannosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D054742:Acyl-Butyrolactones; D001426:Bacterial Proteins; D044902:beta-Mannosidase