Construction of a sensitive and specific lead biosensor using a genetically engineered bacterial system with a luciferase gene reporter controlled by pbr and cadA promoters

Abstract

Background A bacterial biosensor refers to genetically engineered bacteria that produce an assessable signal in the presence of a physical or chemical agent in the environment. Methods We have designed and evaluated a bacterial biosensor expressing a luciferase reporter gene controlled by pbr and cadA promoters in Cupriavidus metallidurans (previously termed Ralstonia metallidurans) containing the CH34 and pI258 plasmids of Staphylococcus aureus, respectively, and that can be used for the detection of heavy metals. In the present study, we have produced and evaluated biosensor plasmids designated pGL3-luc/pbr biosensor and pGL3-luc/cad biosensor, that were based on the expression of luc+ and under the control of the cad promoter and the cadC gene of S. aureus plasmid pI258 and pbr promoter and pbrR gene from plasmid pMOL30 of Cupriavidus metallidurans. Results We found that the pGL3-luc/pbr biosensor may be used to measure lead concentrations between 1–100 μM in the presence of other metals, including zinc, cadmium, tin and nickel. The latter metals did not result in any significant signal. The pGL3-luc/cad biosensor could detect lead concentrations between 10 nM to 10 μM. Conclusions This biosensor was found to be specific for measuring lead ions in both environmental and biological samples.

Transcription factor-based biosensors for screening and dynamic regulation

“…One should consider, among other factors, the excitation and emission wavelength, the maturation time ( Shaner et al, 2005 ) and the half-life of the matured protein [as the FP can sometimes be too stable and rendered useless in real-time applications such as biosensing ( Andersen et al, 1998 )]. Bioluminescent proteins are other alternative reporters ( Nourmohammadi et al, 2020 ; Hansen et al, 2021 ) that rely on biochemical reactions emitting photons as products. Compared to fluorescent proteins, bioluminescent reporters do not rely on the measuring equipment to excite a fluorophore, which leads to less background emission and higher sensitivity.…”

Modern analytical and bioanalytical technologies and concepts for smart and precision farming

“…The specicity and selectivity of biosensors toward toxic heavy metals, including Pb 2+ , Hg 2+ , and As 3+ , can be further improved by inserting recombinant plasmids with specic promoters that trigger specic enzyme reactions, such as activation of the luciferase enzyme and b-galactosidase. 15,16 A whole-cell biosensor is an alternative to an enzyme biosensor, as it is relatively cheap to fabricate, as the isolation step for biomolecules is eliminated. Other advantages include high stability and the ability to respond to a broader range of substrates.…”

“…Numerous studies have been conducted on enhancing the performance of biosensors by investigating materials science, such as nanomaterials and polymers, immobilization methods such as adsorption, affinity binding, covalent binding, crosslinking, and entrapment, 13 and the use of biotechnology. [14][15][16] Various efforts have been implemented to overcome challenges in the mass production of biosensors by simplifying and reducing the cost of the fabrication process. [17][18][19][20][21] This review will examine how biosensors have evolved and been used in smart and precise farming over the past ve years.…”

Biosensor Constructs for the Monitoring of Persistent Emerging Pollutants in Environmental Matrices

“…Similar strategies have been followed to design biosensors for cadmium with a LOD of 25 nM and 1.3 μM, 49,65 arsenic with a LOD of 0.1 μM, 66 and lead with a LOD of 10 nM. 67 Furthermore, other microorganisms have been used, such as R. eutropha AE2515 for nickel and cobalt detection with a LOD of 0.1 and 9 μM, respectively, 68 Pseudomonas fluorescens DF57 for copper detection with a LOD of 0.3 ppm, 69 and S. cerevisiae to detect copper with a LOD of 0.5 mM. 50 The detection of aromatic compounds such as phenols, benzene, and toluene has been also addressed by cell-based biosensors.…”

“…coli. ,, The limit of detection (LOD) of these mercury biosensors varies in the range of concentrations from nanomolar to picomolar. Similar strategies have been followed to design biosensors for cadmium with a LOD of 25 nM and 1.3 μM, , arsenic with a LOD of 0.1 μM, and lead with a LOD of 10 nM . Furthermore, other microorganisms have been used, such as R.…”