Engineering Rugged Field Assays to Detect Hazardous Chemicals Using Spore-Based Bacterial Biosensors

Wynn, Daniel T.; Deo, Sapna K.; Daunert, Sylvia

doi:10.1016/bs.mie.2017.02.005

Cited by 10 publications

(10 citation statements)

References 111 publications

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“…Storing the biosensor strains as spores enables both long-term storage and transport without the need of special cooling systems [41]. Upon arrival at the designated operation site, the cells can easily be revived from the spores and are ready to use within a few hours without any loss in performance, based on the experience with other B. subtilis biosensors [42,43]. Previous studies have already demonstrated the advantage of using spores as a storage system for whole-cell biosensors, thereby extending the life span and making them withstand unfavourable environmental conditions [42].…”

Section: Discussionmentioning

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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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%

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

Lautenschläger

Mascher

2020

J Biol Eng

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“…Structurally, biosensors are a complex consisting of three main functional elements: (I) bioreceptor (bio-recognition component) with the elements for recognizing target analytes (target molecules) contained in biosubstrates, which are located on the sensor plate; (II) transducer, acting based on physicochemical (electrochemical, spectroscopic, or optical) principles; and (III) electronic device for signal processing, recording, and displaying data in a form (analog or digital) convenient for the researcher [25,[33][34][35][36] (Figure 2). Over the decades of development of the biosensing technology, a large number of structurally different sensors and approaches to systematize them have been proposed [25,29,[37][38][39][40].…”

Section: Main Types Of Biosensors and Their Functionsmentioning

confidence: 99%

Label-Free Biosensors for Laboratory-Based Diagnostics of Infections: Current Achievements and New Trends

et al. 2020

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Infections pose a serious global public health problem and are a major cause of premature mortality worldwide. One of the most challenging objectives faced by modern medicine is timely and accurate laboratory-based diagnostics of infectious diseases. Being a key factor of timely initiation and success of treatment, it may potentially provide reduction in incidence of a disease, as well as prevent outbreak and spread of dangerous epidemics. The traditional methods of laboratory-based diagnostics of infectious diseases are quite time- and labor-consuming, require expensive equipment and qualified personnel, which restricts their use in case of limited resources. Over the past six decades, diagnostic technologies based on lateral flow immunoassay (LFIA) have been and remain true alternatives to modern laboratory analyzers and have been successfully used to quickly detect molecular ligands in biosubstrates to diagnose many infectious diseases and septic conditions. These devices are considered as simplified formats of modern biosensors. Recent advances in the development of label-free biosensor technologies have made them promising diagnostic tools that combine rapid pathogen indication, simplicity, user-friendliness, operational efficiency, accuracy, and cost effectiveness, with a trend towards creation of portable platforms. These qualities exceed the generally accepted standards of microbiological and immunological diagnostics and open up a broad range of applications of these analytical systems in clinical practice immediately at the site of medical care (point-of-care concept, POC). A great variety of modern nanoarchitectonics of biosensors are based on the use of a broad range of analytical and constructive strategies and identification of various regulatory and functional molecular markers associated with infectious bacterial pathogens. Resolution of the existing biosensing issues will provide rapid development of diagnostic biotechnologies.

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“…Whole cell biosensors for checking the presence, detection and biodegradation potential of xenobiotics compounds (pharmaceutical residues, pesticides, paraffin, PAHs and PCBs, etc.) present (Adhikari, 2019) in environmental samples are attaining attention (Wynn et al, 2017;Heng et al, 2018;Patel et al, 2019). The reporter proteins acting microbe makes a color signal at the detection of particular contaminants via transducer (Zhang and Liu, 2016).…”

Section: Synthetic Genetic Circuit and Microbial Biosensormentioning

confidence: 99%

Alternative Strategies for Microbial Remediation of Pollutants via Synthetic Biology

2020

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Continuous contamination of the environment with xenobiotics and related recalcitrant compounds has emerged as a serious pollution threat. Bioremediation is the key to eliminating persistent contaminants from the environment. Traditional bioremediation processes show limitations, therefore it is necessary to discover new bioremediation technologies for better results. In this review we provide an outlook of alternative strategies for bioremediation via synthetic biology, including exploring the prerequisites for analysis of research data for developing synthetic biological models of microbial bioremediation. Moreover, cell coordination in synthetic microbial community, cell signaling, and quorum sensing as engineered for enhanced bioremediation strategies are described, along with promising gene editing tools for obtaining the host with target gene sequences responsible for the degradation of recalcitrant compounds. The synthetic genetic circuit and two-component regulatory system (TCRS)-based microbial biosensors for detection and bioremediation are also briefly explained. These developments are expected to increase the efficiency of bioremediation strategies for best results.

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Engineering Rugged Field Assays to Detect Hazardous Chemicals Using Spore-Based Bacterial Biosensors

Cited by 10 publications

References 111 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

Label-Free Biosensors for Laboratory-Based Diagnostics of Infections: Current Achievements and New Trends

Alternative Strategies for Microbial Remediation of Pollutants via Synthetic Biology

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