Development of a whole-cell biosensor for detection of antibiotics targeting bacterial cell envelope in Bacillus subtilis

2022 ◽  
Author(s):  
Jianhua Yin ◽  
Dan Cheng ◽  
Yiling Zhu ◽  
Yanqun Liang ◽  
Zhiliang Yu
Keyword(s):  
Bacillus Subtilis ◽  
Bacterial Cell ◽  
Cell Envelope ◽  
Whole Cell ◽  
Bacterial Cell Envelope ◽  
Whole Cell Biosensor ◽  
Cell Biosensor

scholarly journals Expression of the Lipopolysaccharide-Modifying Enzymes PagP and PagL Modulates the Endotoxic Activity of Bordetella pertussis

2006 ◽  
Vol 74 (10) ◽  
pp. 5574-5585 ◽  
Author(s):  
Jeroen Geurtsen ◽  
Liana Steeghs ◽  
Hendrik-Jan Hamstra ◽  
Jan ten Hove ◽  
Alex de Haan ◽  
...  
Keyword(s):  
Pertussis Vaccine ◽  
Bordetella Pertussis ◽  
Bacterial Cell ◽  
Cell Envelope ◽  
Bacterial Cells ◽  
Gram Negative ◽  
Whole Cell ◽  
Bacterial Cell Envelope ◽  
Paradoxical Result ◽  
Endotoxic Activity

ABSTRACT Lipopolysaccharide (LPS) is one of the major constituents of the gram-negative bacterial cell envelope. Its endotoxic activity causes the relatively high reactogenicity of whole-cell vaccines. Several bacteria harbor LPS-modifying enzymes that modulate the endotoxic activity of the LPS. Here we evaluated whether two such enzymes, i.e., PagP and PagL, could be useful tools for the development of an improved and less reactogenic whole-cell pertussis vaccine. We showed that expression of PagP and PagL in Bordetella pertussis leads to increased and decreased endotoxic activity of the LPS, respectively. As expected, PagP activity also resulted in increased endotoxic activity of whole bacterial cells. However, more unexpectedly, this was also the case for PagL. This paradoxical result may be explained, in part, by an increased release of LPS, which we observed in the PagL-expressing cells.

scholarly journals User experience: Using national Cryo EM centers towards studying lipid transport across the bacterial cell envelope

2021 ◽  
Vol 27 (S1) ◽  
pp. 1422-1422
Author(s):  
Gira Bhabha ◽  
Damian Ekiert ◽  
Nicolas Coudray ◽  
Georgia Isom ◽  
Mark MacRae ◽  
...  
Keyword(s):  
User Experience ◽  
Bacterial Cell ◽  
Cell Envelope ◽  
Lipid Transport ◽  
Bacterial Cell Envelope

scholarly journals Influence of the luxR Regulatory Gene Dosage and Expression Level on the Sensitivity of the Whole-Cell Biosensor to Acyl-Homoserine Lactone

Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 166
Author(s):  
Sergey Bazhenov ◽  
Uliana Novoyatlova ◽  
Ekaterina Scheglova ◽  
Vadim Fomin ◽  
Svetlana Khrulnova ◽  
...  
Keyword(s):  
Gene Dosage ◽  
Regulatory Gene ◽  
Homoserine Lactone ◽  
Threshold Concentration ◽  
Expression Level ◽  
Whole Cell ◽  
Lux Biosensors ◽  
Order Of Magnitude ◽  
Whole Cell Biosensor ◽  
Cell Biosensor

Aliivibrio fischeri LuxR and Aliivibrio logei LuxR1 and LuxR2 regulatory proteins are quorum sensing transcriptional (QS) activators, inducing promoters of luxICDABEG genes in the presence of an autoinducer (3-oxo-hexanoyl-l-homoserine lactone). In the Aliivibrio cells, luxR genes are regulated by HNS, CRP, LitR, etc. Here we investigated the role of the luxR expression level in LuxI/R QS system functionality and improved the whole-cell biosensor for autoinducer detection. Escherichia coli-based bacterial lux-biosensors were used, in which Photorhabdus luminescensluxCDABE genes were controlled by LuxR-dependent promoters and luxR, luxR1, or luxR2 regulatory genes. We varied either the dosage of the regulatory gene in the cells using additional plasmids, or the level of the regulatory gene expression using the lactose operon promoter. It was shown that an increase in expression level, as well as dosage of the regulatory gene in biosensor cells, leads to an increase in sensitivity (the threshold concentration of AI is reduced by one order of magnitude) and to a two to threefold reduction in response time. The best parameters were obtained for a biosensor with an increased dosage of luxRA. fischeri (sensitivity to 3-oxo-hexanoyl-l-homoserine lactone reached 30–100 pM).

scholarly journals Detection of Bioavailable Cadmium by Double-Color Fluorescence Based on a Dual-Sensing Bioreporter System

2021 ◽  
Vol 12 ◽  
Author(s):  
Chang-ye Hui ◽  
Yan Guo ◽  
Jian Wu ◽  
Lisa Liu ◽  
Xue-qin Yang ◽  
...  
Keyword(s):  
Anthropogenic Activities ◽  
Simultaneous Detection ◽  
Environmental Water ◽  
High Concentration ◽  
Whole Cell ◽  
Specificity And Sensitivity ◽  
Improved Performance ◽  
Whole Cell Biosensor ◽  
Cell Biosensor ◽  
Dual Sensing

Cadmium (Cd) is carcinogenic to humans and can accumulate in the liver, kidneys, and bones. There is widespread presence of cadmium in the environment as a consequence of anthropogenic activities. It is important to detect cadmium in the environment to prevent further exposure to humans. Previous whole-cell biosensor designs were focused on single-sensing constructs but have had difficulty in distinguishing cadmium from other metal ions such as lead (Pb) and mercury (Hg). We developed a dual-sensing bacterial bioreporter system to detect bioavailable cadmium by employing CadC and CadR as separate metal sensory elements and eGFP and mCherry as fluorescent reporters in one genetic construct. The capability of this dual-sensing biosensor was proved to simultaneously detect bioavailable cadmium and its toxic effects using two sets of sensing systems while still maintaining similar specificity and sensitivity of respective signal-sensing biosensors. The productions of double-color fluorescence were directly proportional to the exposure concentration of cadmium, thereby serving as an effective quantitative biosensor to detect bioavailable cadmium. This novel dual-sensing biosensor was then validated to respond to Cd(II) spiked in environmental water samples. This is the first report of the development of a novel dual-sensing, whole-cell biosensor for simultaneous detection of bioavailable cadmium. The application of two biosensing modules provides versatile biosensing signals and improved performance that can make a significant impact on monitoring high concentration of bioavailable Cd(II) in environmental water to reduce human exposure to the harmful effects of cadmium.

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