Novel Cyanobacterial Biosensor for Detection of Herbicides

Abstract

The aim of this work was to generate a cyanobacterial biosensor that could be used to detect herbicides and other environmental pollutants. A representative freshwater cyanobacterium, Synechocystis sp. strain PCC6803, was chromosomally marked with the luciferase gene luc (from the firefly Photinus pyralis) to create a novel bioluminescent cyanobacterial strain. Successful expression of the luc gene during growth of Synechocystis sp. strain PCC6803 cultures was characterized by measuring optical density and bioluminescence. Bioluminescence was optimized with regard to uptake of the luciferase substrate, luciferin, and the physiology of the cyanobacterium. Bioassays demonstrated that a novel luminescent cyanobacterial biosensor has been developed which responded to a range of compounds including different herbicide types and other toxins. This biosensor is expected to provide new opportunities for the rapid screening of environmental samples or for the investigation of potential environmental damage.

A New, Sensitive Marine Microalgal Recombinant Biosensor Using Luminescence Monitoring for Toxicity Testing of Antifouling Biocides

“…An alternative strategy is to develop genetic engineering tools to express recombinant luciferase (Luc) in fastgrowing microorganisms expressing recombinant luciferase. Recombinant microorganisms, such as Escherichia coli encoding the thermostable lux luciferase from Photorhabdus luminescens (12) and Synechocystis sp., a freshwater cyanobacterium, encoding the thermostable luciferase from the firefly Photinus pyralis (13) have been used for both industrial effluent (14,15) and soil environment (16,17) monitoring. To the best of our knowledge, marine microorganism recombinant biosensors have never been developed for ecotoxicological testing.…”

Microbiological Biosensors Based on Luminescence

“…Biorąc pod uwagę najnowszą literaturę, największą popularność odnotowano w ochronie środowiska, gdzie za pomocą mikrobiologicznych biosensorów luminescencyjnych wykonuje się: pomiar toksyczności wody [22,24,25], analizę pod kątem zawartości w niej szkodliwych jonów Zn, Cu, Cd [18] czy zawartości arsenu [26]. Mikrobiologiczne sensory luminescencyjne wykorzystywane są również w przemyśle spożywczym, gdzie używane są do monitorowania zawartości szkodliwych bakterii w pożywieniu [27].…”

Microbial Cell Arrays

“…The latter category includes bacterial luciferase (lux) and b-galactosidase (lacZ), as well as others: insect luciferase (luc), alkaline phosphatase (phoA), b-glucuronidase (uidA), and b-lactamase (bla) [63][64][65][66][67]. Depending on the reporter gene used and on the substrate provided to its product, the emitted signals can be detected optically (by bioluminescence, fluorescence, or colorimetry), or electrochemically [57].…”