Selective inhibition of Zophobas morio (Coleoptera: Tenebrionidae) luciferase-like enzyme luminescence by diclofenac and potential suitability for light-off biosensing

Autor(es)

Carvalho, Mariele C. ^[1] ; Tomazini, Atilio ^[2] ; Prado, Rogilene A. ^[3] ; Viviani, Vadim R. ^{[1, 3]}

Afiliação do(s) autor(es)

^[1] Fed Univ Sao Carlos UFSCar, Grad Program Evolut Genet & Mol Biol, Sao Carlos - Brazil

^[2] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Biorenewables Natl Lab LNBR, Campinas, SP - Brazil

^[3] Fed Univ Sao Carlos UFSCar, Grad Program Biotechnol & Environm Monitoring, Sorocaba - Brazil

• Tipo de documento: Artigo Científico
• Fonte: LUMINESCENCE; v. 36, n. 2 NOV 2020.
• Citações Web of Science: 0

Resumo

The accumulation of toxic carboxylic compounds may cause severe effects on the environment and living organisms. A luciferase-like enzyme, previously cloned from the Malpighian tubules of the non-luminescent Zophobas morio mealworm, displays thioesterification activity with a wide range of carboxylic substrates, and produces weak red luminescence in the presence of ATP and firefly d-luciferin, a xenobiotic for this organism. To better investigate the function of this enzyme in carboxylic xenobiotic detoxification, we analyzed the inhibitory effect of different xenobiotic carboxylic acids on the luminescence activity of this enzyme, including environmental pollutants and pharmaceutical compounds. Noteworthy, the anti-inflammatory drug diclofenac severely inhibited this luciferase-like enzyme luminescence activity, both in in vitro (IC50 20 mu M) and in vivo in bacterial cells assays, when compared with other beetle luciferases. Similar results were obtained with its brighter I327S mutant. Kinetic analysis of diclofenac's effect on luminescence activity indicated mixed-type inhibition for both ATP and d-luciferin. Modelling studies showed five potential binding sites for diclofenac, including the coenzyme A binding site, which showed one of the highest binding constant. Taken together, these results raise the possibility of using this luciferase-like enzyme for the development of novel whole-cell luminescent biosensors for diclofenac and similar drugs. (AU)

• Processo FAPESP: 16/07312-6 - Estrutura e função biológica da enzima tipo-luciferase de lanternas de Arachnocampa luminosa (Diptera: Keroplatidae) e desenvolvimento de atividade luminescente por Engenharia genética
• Beneficiário: Mariele Cristina de Carvalho
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 14/18179-0 - Emprego da enzima tipo-luciferase de Zophobas morio recombinante para bioensaios de toxicidade de compostos carboxílicos xenobióticos
• Beneficiário: Mariele Cristina de Carvalho
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 10/05426-8 - Bioluminescência de artrópodes: diversidade biológica em biomas brasileiros; origem bioquímica; evolução estrutural/funcional de luciferases; diferenciação molecular das lanternas; aplicações biotecnológicas, ambientais e educacionais
• Beneficiário: Vadim Viviani
• Modalidade de apoio: Auxílio à Pesquisa - Temático