title: Development of a Pseudomonas aeruginosa Agmatine Biosensor.
abstract: Agmatine, decarboxylated arginine, is an important intermediary in polyamine production for many prokaryotes, but serves higher functions in eukaryotes such as nitric oxide inhibition and roles in neurotransmission. Pseudomonas aeruginosa relies on the arginine decarboxylase and agmatine deiminase pathways to convert arginine into putrescine. One of the two known agmatine deiminase operons, aguBA, contains an agmatine sensitive TetR promoter controlled by AguR. We have discovered that this promoter element can produce a titratable induction of its gene products in response to agmatine, and utilized this discovery to make a luminescent agmatine biosensor in P. aeruginosa. The genome of the P. aeruginosa lab strain UCBPP-PA14 was altered to remove both its ability to synthesize or destroy agmatine, and insertion of the luminescent reporter construct allows it to produce light in proportion to the amount of exogenous agmatine applied from ~100 nM to 1mM. Furthermore it does not respond to related compounds including arginine or putrescine. To demonstrate potential applications the biosensor was used to detect agmatine in spent supernatants, to monitor the development of arginine decarboxylase over time, and to detect agmatine in the spinal cords of live mice.
authors.affiliation: Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, University of Minnesota, 420 Delaware St. SE MMC 276, Minneapolis, MN 55455, USA; E-Mail: gilbe398@umn.edu.
authors.forename: Adam
authors.initials: A
authors.lastname: Gilbertsen
authors.affiliation: Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, University of Minnesota, 420 Delaware St. SE MMC 276, Minneapolis, MN 55455, USA; E-Mail: gilbe398@umn.edu.
authors.forename: Bryan
authors.initials: B
authors.lastname: Williams
keywords: Pseudomonas; agmatine; arginine decarboxylase; bioluminescence; polyamines