Photoproteins as Reporters in Whole‐cell Sensing

Feliciano, Jessika; Pasini, Patrizia; Deo, Sapna K.; Daunert, Sylvia

doi:10.1002/3527609148.ch8

Cited by 8 publications

(4 citation statements)

References 120 publications

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“…Biosensors of this type have been developed for a variety of analytes. More in-depth information and examples can be found in a number of reviews. − Specifically, we have developed a whole-cell sensing system for the detection of OH-PCBs by employing the strain Pseudomonas azelaica HBP1. This bacterium contains the hbpCAD genes, which are responsible for the degradation of hydroxylated biphenyls.…”

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confidence: 99%

Hydroxylated Polychlorinated Biphenyl Detection Based on a Genetically Engineered Bioluminescent Whole-Cell Sensing System

Turner

Pasini

et al. 2007

Anal. Chem.

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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.

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Hydroxylated Polychlorinated Biphenyl Detection Based on a Genetically Engineered Bioluminescent Whole-Cell Sensing System

Turner

Pasini

et al. 2007

Anal. Chem.

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“…Whole-cell sensing systems possess all the characteristics required for the development of such methods. These sensing methods have found applications in biotechnology, pharmacology, molecular biology, and environmental as well as clinical chemistry. − In genetically engineered whole-cell biosensors, genetic constructs are employed in which a regulatory protein capable of molecular recognition is designed to control the expression of a reporter protein. When the target analyte is present, the regulatory protein recognizes it and activates gene transcription.…”

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Biosensing Systems for the Detection of Bacterial Quorum Signaling Molecules

et al. 2006

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Bacterial quorum sensing (QS) is a cell-to-cell communication phenomenon that allows bacteria to control the expression of certain specialized genes depending on their cell population size. Signaling molecules such N-acylhomoserine lactones (AHLs) mediate the communication, and their concentration reflects the bacterial population density. Quorum sensing regulates several processes including bacterial pathogenicity. We developed a method for the rapid, sensitive, and quantitative detection of AHLs in biological samples such as saliva and stools. The method is based on whole-cell sensing systems that employ QS regulatory systems as recognition elements and the luxCDABE gene cassette as a reporter. The method proved to be reproducible when applied to real samples and was able to detect low analyte concentrations down to 1 x 10(-9) M without requiring extensive sample preparation. We envision that this novel biosensing system could be employed in the diagnosis and management of various bacteria-related disorders, thus supporting the use of quorum sensing molecules as potential biomarkers of disease. Due to cost-effectiveness and high throughput, these biosensing systems could be successfully employed as a new tool for the screening of novel drugs that target quorum sensing mechanisms.

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“…Whole cell biosensing systems have several features that render them suitable components of portable field kits. These characteristics include ability to withstand environmental conditions, such as a wide ranges of temperatures, pHs, and ionic strengths; provide information on the analyte bioavailability; requirement of minimal or no sample pretreatment; high sensitivity and selectivity; ease of preparation; easy and rapid detection; cost-effectiveness; amenability to high-throughput screening; miniaturization; and automation. , …”

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Paper Strip Whole Cell Biosensors: A Portable Test for the Semiquantitative Detection of Bacterial Quorum Signaling Molecules

et al. 2010

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Herein, we report the development of a novel, inexpensive, and portable filter-paper-based strip biosensor for the detection of bacterial quorum sensing signaling molecules, N-acylhomoserine lactones (AHLs). AHLs are generally employed by Gram-negative bacteria for their cell-cell communication to control expression of specialized genes, such as those involved in biofilm formation and production of virulence factors, in a population-density-dependent manner. First, a bacterial cell-based sensing system employing components of AHL-mediated QS regulatory system as recognition elements and beta-galactosidase as the reporter protein was designed and developed. The bacterial-sensing cells were then liquid-dried on strips of filter paper. beta-Galactosidase as the reporter allows for the visual monitoring of the analyte-induced signal when a colorimetric method of detection is applied. The paper strip biosensor was able to detect low AHL concentrations down to 1 x 10(-8) M. Furthermore, it was successfully applied to the detection of AHLs in physiological samples, such as saliva. The filter-paper-based sensing strips could provide reproducible results upon storage at 4 degrees C for at least 3 months. In conclusion, a filter-paper-based strip biosensor was developed that allows for visual, fast, and convenient detection of AHLs in a dose-dependent manner in a test sample. In addition, it does not require expensive equipment or trained personnel and allows ease of transportation and storage. Therefore, we envision that this biosensor will serve as a simple and economical portable field kit for on-site monitoring of AHL in a variety of clinical and environmental samples.

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Photoproteins as Reporters in Whole‐cell Sensing

Cited by 8 publications

References 120 publications

Hydroxylated Polychlorinated Biphenyl Detection Based on a Genetically Engineered Bioluminescent Whole-Cell Sensing System

Hydroxylated Polychlorinated Biphenyl Detection Based on a Genetically Engineered Bioluminescent Whole-Cell Sensing System

Biosensing Systems for the Detection of Bacterial Quorum Signaling Molecules

Paper Strip Whole Cell Biosensors: A Portable Test for the Semiquantitative Detection of Bacterial Quorum Signaling Molecules

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