Recently we cloned an AMP-CoA-ligase with weak luminescence activity in the presence of D-luciferin from the Malpighian tubules of the non-luminescenct Zophobas morio (Coleóptera: Tenebrionidae). The presence of endoplasmic reticulum signal, the ocurrence of the enzyme in the Malpighian tubules,the ligasic and oxidative reactions with the xenbiotic D-luciferin, and the promiscuous activity with several carboxylic substrates, lead us to propose that the enzyme could be involved with the detoxification/excretion of xenobiotics. The enzyme and its properties are object of a patent application at INPI. We aim to investigate the involvement of this enzyme in detoxification and excretion of xenobiotic carboxylic acids in Escherichia coli, and to establish a simple bioassay for toxic carboxylic acids based on the inhibition of luminescence and cellular growth, for later development of a biosensor. We will analyze the growth and cell viability in bacteria transformed with the gene coding the luciferase-like enzyme in the presence of toxic carboxylic acids, and as a control bacteria transformed only with the plasmid without the gene for the luciferase-like enzyme, and assay the luminescence in presence and absence of the acids. The growth inhibition and viability assays in presence of acids will be used as parameters to indicate toxicity, whereas the luminescence and growth of transformed cells will be used as parameters to indicate the detoxification of these compounds.
News published in Agência FAPESP Newsletter about the scholarship: