Sposensor: A Whole-Bacterial Biosensor That Uses Immobilized &lt;i&gt;Bacillus subtilis&lt;/i&gt; Spores and a One-Step Incubation/Detection Process

Fantino, Jean-Raphaël; Barras, Frédéric; Denizot, François

doi:10.1159/000206634

Cited by 17 publications

(13 citation statements)

References 48 publications

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“…Since our whole-cell biosensor is based on B. subtilis, it is potentially also adaptable for applications outside of the laboratory and in low-tech environments. By replacing the luciferase reporter with the βgalactosidase reporter, the induction becomes visible by eye [18,19]. Storing the biosensor strains as spores enables both long-term storage and transport without the need of special cooling systems [41].…”

Section: Discussionmentioning

confidence: 99%

“…The value of applying biosensors is their ease of use and the low costs compared to chromatography-based detection methods or immunoassays that require expensive equipment or experienced staff [16,17]. Freeze-drying or the use of bacterial spores for transport allow using biosensors in the field, where they can be 'revived' by rehydration at the designated operation site [18,19]. While a defined specificity for a certain class of compounds is a prerequisite for a good biosensor, achieving the necessary sensitivity to detect low compound concentrations can be challenging [17].…”

Section: Introductionmentioning

confidence: 99%

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Development of a novel heterologous β-lactam-specific whole-cell biosensor in Bacillus subtilis

Lautenschläger

Mascher

2020

J Biol Eng

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Background: Whole-cell biosensors are a powerful and easy-to-use screening tool for the fast and sensitive detection of chemical compounds, such as antibiotics. β-Lactams still represent one of the most important antibiotic groups in therapeutic use. They interfere with late stages of the bacterial cell wall biosynthesis and result in irreversible perturbations of cell division and growth, ultimately leading to cell lysis. In order to simplify the detection of these antibiotics from solutions, solid media or directly from producing organisms, we aimed at developing a novel heterologous whole-cell biosensor in Bacillus subtilis, based on the β-lactam-induced regulatory system BlaR1/BlaI from Staphylococcus aureus. Results: The BlaR1/BlaI system was heterologously expressed in B. subtilis and combined with the luxABCDE operon of Photorhabdus luminescens under control of the BlaR1/BlaI target promoter to measure the output of the biosensor. A combination of codon adaptation, constitutive expression of blaR1 and blaI and the allelic replacement of penP increased the inducer spectrum and dynamic range of the biosensor. β-Lactams from all four classes induced the target promoter P blaZ in a concentration-dependent manner, with a dynamic range of 7-to 53-fold. We applied our biosensor to a set of Streptomycetes soil isolates and demonstrated its potential to screen for the production of β-lactams. In addition to the successful implementation of a highly sensitive β-lactam biosensor, our results also provide the first experimental evidence to support previous suggestions that PenP functions as a βlactamase in B. subtilis. Conclusion: We have successfully established a novel heterologous whole-cell biosensor in B. subtilis that is highly sensitive for a broad spectrum of β-lactams from all four chemical classes. Therefore, it increases the detectable spectrum of compounds with respect to previous biosensor designs. Our biosensor can readily be applied for identifying β-lactams in liquid or on solid media, as well as for identifying potential β-lactam producers.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of a novel heterologous β-lactam-specific whole-cell biosensor in Bacillus subtilis

Lautenschläger

Mascher

2020

J Biol Eng

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“…More specifically, as a good example, E. coli cell surface display technology has been applied to construct biosensors [69]. In addition, B. subtilis spores also have been used to construct biosensors [70]. However, to the best of our knowledge, there are no studies on the application of BSSD technology to biosensors.…”

Section: Future Outlookmentioning

confidence: 99%

Bacillus subtilis Spore Surface Display Technology: A Review of Its Development and Applications

Zhang

Zabed

et al. 2019

Journal of Microbiology and Biotechnology

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Bacillus subtilis spore surface display (BSSD) technology is considered to be one of the most promising approaches for expressing heterologous proteins with high activity and stability. Currently, this technology is used for various purposes, such as the production of enzymes, oral vaccines, drugs and multimeric proteins, and the control of environmental pollution. This paper presents an overview of the latest developments in BSSD technology and its application in protein engineering. Finally, the major limitations of this technology and future directions for its research are discussed.

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“…The reporter proteins encoded by such genes have also been used as signal-transduction elements in bacterial sensors. These proteins include β-galactosidase [11,12], bacterial luciferase [13], firefly luciferase [14], and the green fluorescent protein and its variants. Table 1 lists reporters that are commonly used in whole-cell sensing systems, with their catalyzed reactions and methods of detection.…”

Section: Bacterial Whole-cell-based Biosensing Systemsmentioning

confidence: 99%

Engineered cells as biosensing systems in biomedical analysis

et al. 2012

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Over the past two decades there have been great advances in biotechnology, including use of nucleic acids, proteins, and whole cells to develop a variety of molecular analytical tools for diagnostic, screening, and pharmaceutical applications. Through manipulation of bacterial plasmids and genomes, bacterial whole-cell sensing systems have been engineered that can serve as novel methods for analyte detection and characterization, and as more efficient and cost-effective alternatives to traditional analytical techniques. Bacterial cell-based sensing systems are typically sensitive, specific and selective, rapid, easy to use, low-cost, and amenable to multiplexing, high-throughput, and miniaturization for incorporation into portable devices. This critical review is intended to provide an overview of available bacterial whole-cell sensing systems for assessment of a variety of clinically relevant analytes. Specifically, we examine whole-cell sensing systems for detection of bacterial quorum sensing molecules, organic and inorganic toxic compounds, and drugs, and for screening of antibacterial compounds for identification of their mechanisms of action. Methods used in the design and development of whole-cell sensing systems are also reviewed.

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Sposensor: A Whole-Bacterial Biosensor That Uses Immobilized <i>Bacillus subtilis</i> Spores and a One-Step Incubation/Detection Process

Cited by 17 publications

References 48 publications

Development of a novel heterologous β-lactam-specific whole-cell biosensor in Bacillus subtilis

Development of a novel heterologous β-lactam-specific whole-cell biosensor in Bacillus subtilis

Bacillus subtilis Spore Surface Display Technology: A Review of Its Development and Applications

Engineered cells as biosensing systems in biomedical analysis

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