Analytical Sciences

Bioluminescence Microplate Assay of Cyanide with Escherichia coli Harboring a Plasmid Responsible for Cyanide-dependent Light Emission in Alginate Microenvironment

Hajime KARATANI,*1,*2,*3 Yasuro FUSE,*1,*3 Hirotaka MIZUGUCHI,*3 Shogo MONJI,*1 Hiroshi OYAMA,*4 Tomonori WAKU,*1 and Masashi IWASAKI*3,*5

*1 Faculty of Molecular Chemistry and Engineering, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo, Kyoto 606-8585, Japan
*2 Kyoto Luminous Science Laboratory, 1 Hashigami-cho, Matsugasaki, Sakyo, Kyoto 606-8585, Japan
*3 Center of Environmental Science, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo, Kyoto 606-8585, Japan
*4 Department of Life Science, Faculty of Science and Engineering, Setsunan University, 17-8 Ikedanaka-machi, Neyagawa, Osaka 572-8508, Japan
*5 Faculty of Materials Science and Engineering, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo, Kyoto 606-8585, Japan

We describe the bioluminescence of a genetically engineered Escherichia coli harboring a recombined plasmid with a catalase gene promoter fused lux gene cluster, responsible for the generation of photons closely associated with respiratory inhibition, with the aim of applying it for cyanide sensing. This E. coli construct was favorably utilized for the microplate assay of cyanide by leveraging the microenvironment of the biocompatible alginate. The brightness of the bioluminescence, induced by cyanide stimulation of the respiration causative of the production of hydrogen peroxide, positively correlates with its concentration. Moreover, visualization of cyanide with a consumer digital camera, ranging in concentration from about 0.01 mg CN·L⁻¹ in the alginate sol to around 100 mg CN·L⁻¹ in its gel, was attained.

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