title: Hydroxylated polychlorinated biphenyl detection based on a genetically engineered bioluminescent whole-cell sensing system.
abstract: The metabolites of polychlorinated biphenyls (PCBs), such as hydroxylated PCBs (OH-PCBs), have been identified as environmental contaminants. Various studies have shown that some OH-PCBs can potentially contribute to health problems. Detection of these compounds in environmental and biological samples could provide useful information about their levels and lead to a better understanding of their apparent toxicity. To that end, we have developed a whole-cell sensing system for the detection of OH-PCBs by taking advantage of the recognition of a group of related compounds, i.e., hydroxylated biphenyls, by the product of the hbpR gene in the hbp operon from Pseudomonas azelaica strain HBP1. By fusing the luxAB genes, encoding the reporter protein bacterial luciferase, to the hbp regulator-promoter sequence, a whole-cell sensing system was developed. Here, we describe the optimization and application of this whole-cell sensing system for the detection of a model compound, 2-hydroxy-3',4'-dichlorobiphenyl. A detection limit of 1 x 10(-8) M was achieved using this system. The detection of a broad range of individual OH-PCBs as well as an OH-PCB mixture was investigated. The system can detect OH-PCBs in whole serum samples in a trace amount, which is comparable to the detection of these analytes in medium alone. We envision that the method developed can potentially be employed as a rapid and sensitive way to monitor OH-PCBs for toxicological study in the laboratory, as well as a useful tool to evaluate the presence of bioavailable OH-PCBs in natural environments.
authors.affiliation: Department of Chemistry, University of Kentucky, Lexington, Kentucky 40506-0055, USA.
authors.forename: Kendrick
authors.initials: K
authors.lastname: Turner
authors.forename: Shifen
authors.initials: S
authors.lastname: Xu
authors.forename: Patrizia
authors.initials: P
authors.lastname: Pasini
authors.forename: Sapna
authors.initials: S
authors.lastname: Deo
authors.forename: Leonidas
authors.initials: L
authors.lastname: Bachas
authors.forename: Sylvia
authors.initials: S
authors.lastname: Daunert
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004785:Environmental Pollutants / Q000032:analysis* / Q000633:toxicity*; D017930:Genes, Reporter / Q000235:genetics / Q000502:physiology; D006801:Humans; D006900:Hydroxylation; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D011078:Polychlorinated Biphenyls / Q000032:analysis* / Q000633:toxicity*; D011506:Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D004785:Environmental Pollutants; D011506:Proteins; D011078:Polychlorinated Biphenyls; D008156:Luciferases