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.
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Acknowledgments
This work was supported by a grant from the National Natural Sciences Foundation of China (u1170303). The authors would like to thank Prof. Qifa Zhang (National Center of Plant Gene Research, PRC) for providing many useful suggestions, Dr. Paul Williams (University of Nottingham, UK) for supplying plasmid pSB403, and Dr. Lianhui Zhang (Institute of Molecular and Cell Biology, Singapore) for providing the sensor strain of A. tumefaciens, strain NT1 (traR; tra::lacZ749).
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Liu, P., Gao, Y., Huang, W. et al. A Novel Bioassay for High-Throughput Screening Microorganisms with N-acyl Homoserine Lactone Degrading Activity. Appl Biochem Biotechnol 167, 73–80 (2012). https://doi.org/10.1007/s12010-012-9653-4
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DOI: https://doi.org/10.1007/s12010-012-9653-4