title: Urate Oxidase produced by Lucilia sericata medical maggots is localized in Malpighian tubes and facilitates allantoin production.
abstract: Lucilia sericata maggots are the only species currently approved for maggot debridement therapy (MDT), an alternative treatment for chronic and recalcitrant wounds. Maggots promote wound debridement, disinfection and healing by producing a complex mixture of proteins, peptides and low-molecular-weight compounds in their secretions and excretions, but the individual components are not well characterized at the molecular level. Here we investigated the purine catabolism pathway in L. sericata, focusing on the production of allantoin by Urate Oxidase (UO), which is thought to promote wound healing. We produced recombinant L. sericata UO in Escherichia coli, and characterized the properties of the pure enzyme in terms of the optimum pH (7-10) and temperature (20-25 °C), its stability, sensitivity to inhibition and ion dependency. We used quantitative RT-PCR and RNA in situ hybridization to monitor the expression of the UO gene, and we used a guinea pig anti-UO antibody to detect the native enzyme by western blot and by florescence immunohistochemistry in larval tissues. We found that L. sericata UO is exclusively present in the larval excretion organ (the Malpighian tubes) and is freely available in the cytoplasm rather than restricted to a specific subcellular compartment. Allantoin is a final product of L. sericata purine catabolism. It is produced by UO in the Malpighian tubes to remove uric acid from the hemolymph and is consequently excreted via the hindgut. Our findings confirm the hypothesis that both actively secreted molecules and excretion products contribute to the beneficial effects of MDT.
authors.affiliation: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany.
authors.forename: Andre
authors.initials: A
authors.lastname: Baumann
authors.affiliation: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany.
authors.forename: Marisa
authors.initials: M
authors.lastname: Skaljac
authors.affiliation: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany.
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Lehmann
authors.affiliation: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany; Justus-Liebig-University of Giessen, Institute for Insect Biotechnology, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Vilcinskas
authors.affiliation: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany. Electronic address: zdenek.franta@ime.fraunhofer.de.
authors.forename: Zdenӗk
authors.initials: Z
authors.lastname: Franta
mesh_terms: D000481:Allantoin / Q000378:metabolism*; D000818:Animals; D004175:Diptera / Q000201:enzymology*; D007814:Larva / Q000201:enzymology; D008317:Malpighian Tubules / Q000378:metabolism*; D014503:Urate Oxidase / Q000378:metabolism*
chemical_list: D000481:Allantoin; D014503:Urate Oxidase
keywords: Allantoin; Lucilia sericata; Maggot debridement therapy; Urate Oxidase; Wound healing