Open AccessIntroduction of quorum sensing elements into bacterial bioreporter circuits enhances explosives’ detection capabilities
Author(s) -
Shpigel Etai,
Nathansohn Shiri,
Glozman Anat,
Rosen Rachel,
Shemer Benjamin,
YagurKroll Sharon,
Elad Tal,
Belkin Shimshon
Publication year - 2022
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.202100134
Subject(s) - bioreporter , bioluminescence , quorum sensing , photorhabdus luminescens , reporter gene , biology , escherichia coli , gene cassette , gene , bioaugmentation , microbiology and biotechnology , luciferase , bacteria , genetics , biochemistry , gene expression , virulence , microorganism , transfection , integron
A possible solution for the standoff detection of buried landmines is based on the use of microbial bioreporters, genetically engineered to emit a remotely detectable optical signal in response to trace amounts of explosives’ signature chemicals, mostly 2,4‐dinitrotoluene (DNT). Previously developed DNT sensor strains were based on the fusion of a DNT‐inducible gene promoter to a reporting element, either a fluorescent protein gene or a bacterial bioluminescence gene cassette. In the present study, a different approach was used: the DNT‐inducible promoter activates, in Escherichia coli , the quorum‐sensing luxI and luxR genes of Aliivibrio fischeri . N‐Acyl homoserine lactone (AHL), synthesized by LuxI, combines with LuxR and activates the bioluminescence reporter genes. The resulting bioreporter displayed a dose‐dependent luminescent signal in the presence of DNT. Performance of the sensor strain was further enhanced by manipulation of the sensing element (combining the E. coli DNT‐inducible azoR and yqjF gene promoters), by replacing the luminescence gene cassette of Photorhabdus luminescens luxCDABE with A. fischeri luxCDABEG , and by introducing two mutations, eutE and ygdD , into the host strain. DNT detection sensitivity of the final bioreporter was over 340‐fold higher than the original construct.