Arthropod bioluminescence: biological diversity in Brazilian biomes, biochemical origin, structural/functional evolution of luciferases, molecular differentiation of lanterns, biotechnological, environmental and educational applications

• Grant number: 10/05426-8
• Support Opportunities: Research Projects - Thematic Grants
• Duration: May 01, 2017 - October 31, 2022
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Vadim Viviani
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil
• Associated researchers: Carl Johnson ; Fábio Camargo Abdalla ; Jörg Kobarg ; Mário Tyago Murakami ; Simone Policena Rosa ; Yoshihiro Ohmiya
• Associated grant(s): 17/22262-8 - Multiusuary equipment conceeded by process: 2010/05426-8-bioimaging CCD system, AP.EMU
• Associated scholarship(s): 22/14071-6 - CHARACTERIZATION OF ZZ-AMYLUC FUSION PROTEIN AND ITS OPTIMIZATION FOR BIOLUMINESCENT IMMUNOASSAY PURPOSES, BP.IC; 21/11544-8 - Molecular biology and protein expression techniques, BP.TT; 21/03901-5 - Luciferases expression in mammalian cells for bioassay purposes and phototoxic effect of luciferins and fluorescent compounds from coleoptera and diptera, BP.PD; + associated scholarships 20/07649-6 - Application of bioluminescence for cell tracking and inflammatory processes: in vitro and in vivo study, BP.PD; 18/07925-3 - Transcriptional comparison of the integument of bioluminescent and non-bioluminescent coleopteran larvae during ecdysis; isolation, expression and characterization of coumarate-ligases and laccases; and relation between pigmentation and bioluminescence, BP.PD; 18/26151-9 - Engineering Amydetes vivianii luciferase for use as bioanalytical reagent and as metal biosensor, BP.IC; 18/17855-2 - Cloning, expression and characterization of Phengodidae luciferases and proteins of bioluminescent system: structural, functional and evolutive aspects, BP.DD; 18/19430-9 - Heterologous expression of luciferases in yeasts: potential applications in cell biosensors and bioluminescent immunoassays, BP.PD; 18/03883-4 - Implementation, maintenance and disclosure of luminescence museum, BP.TT; 17/26149-1 - Comparison the photogenic and non-photogenic tissues of bioluminescent keroplatid: morphological and biochemical aspects, BP.IC; 18/02538-1 - Resolution of the three-dimensional structures of beetle luciferases and related enzymes by X-ray crystallography: relationship between structure, bioluminescence spectra and oxygenase activity, BP.PD - associated scholarships

Abstract

Bioluminescence, the emission of cold light by living organisms, has been the subject of many multidisciplinary studies involving physics, chemistry and biology, resulting in important scientific discoveries and biotechnological applications in the last century. It can be considered as one of the signatures of life, and for this reason has been used as an excellent bioindicator at the molecular, cellular, organismic and biospheric levels. In the terrestrial environment it occurs among bacteria, fungi, annelids, mollusks, and especially among the arthropods. Among the insects, the bioluminescent beetles system is well known, whereas the bioluminescent Diptera systems remain largely unknown. Brazil hosts the greatest biodiversity of bioluminescent organisms in the world, including beetles, fungi and marine species. However, with the exception of surveys and studies of bioluminescent beetles Atlantic rain forest, and more recently of fungi, little is known about the bioluminescent biodiversity of the Cerrado biome and Amazon forest which are severely threatened by agricultural progress. In the last decade the cDNAs encoding various luciferases from Brazilian beetles with different properties of bioluminescence (colors, kinetic, pH-sensitivity, thermal stability) were cloned, expressed, purified and investigated under the structural/functional and evolutionary aspects by our research group, revealing important determinants of bioluminescence spectra, pH-sensitivity and oxigenase activity. The resultant knowledge is already allowing to engineer these proteins to improve their biotechnological applicability, and some of these luciferases have already acquired effective commercial application. However, several questions remain open regarding bioluminescent systems of insects, including: (1) the origin and structural/functional evolution of luciferases of Beetles and Diptera; (2) the identity of the luciferases, luciferins and accessory proteins in bioluminescent Diptera; (3) the biochemical and anatomical origins of the bioluminescent apparatus in diptera and Beetles, and their possible evolutionary relationships. In addition, despite the undoubted utility of beetles luciferases and other organisms for bioanalytical purposes, in Brazil we still do not use the unique broad repertoire of recombinant luciferases derived from our fauna for bioanalytical purposes, using imported reagents. Thus, this thematic proposal seeks to achieve broad multidisciplinary study of the terrestrial bioluminescence (especially of insects in Brazil), involving: (i) survey of the biodiversity of bioluminescent arthropods in Cerrado biomes, Amazon Forest, with the investigation of their bioluminescence properties, molecular evolution and phylogeny, and application as environmental bio-indicator; (II) to investigate the bioluminescent system of Diptera, especially the molecular identity of luciferases, luciferins and accessory protein Orfelia fultonii of the Palearctic region; (III) clone and investigate the structure, function and molecular evolution of luciferases and accessory proteins of bioluminescent beetles coming from our fauna and Diptera; (IV) to investigate the anatomical and metabolic origin of bioluminescence in the three families of bioluminescent beetles and Diptera through an approach involving cell biology and functional genomics, and (V) to investigate the applicability of the investigated and optimized luciferases by our research group in the development of biotech, environmental and educational applications (enzyme and cell luminescent biosensors, luminescent bioassays and bio-imaging, educational kits). (AU)

Scientific publications

(29)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MOREIRA, A. C.; AMARAL, D. T.; GABRIEL, G. V. M.; VIVIANI, V. R.. Cloning and molecular properties of a novel luciferase from the Brazilian Bicellonycha lividipennis (Lampyridae: Photurinae) firefly: comparison with other firefly luciferases. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. N/A, p. 13-pg., 2022-05-19. (10/05426-8)

PELENTIR, G. F.; BEVILAQUA, V. R.; VIVIANI, V. R.. A highly efficient, thermostable and cadmium selective firefly luciferase suitable for ratiometric metal and pH biosensing and for sensitive ATP assays. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 18, n. 8, p. 2061-2070, AUG 1 2019. (10/05426-8, 15/06875-4, 18/26151-9)

SILVA, JAQUELINE R.; AMARAL, DANILO T.; VIVIANI, VADIM R.. Comparison of the Malpighian tubules and fat body transcriptional profiles of Zophobas morio larvae (Coleoptera: Tenebrionidae). Comparative Biochemistry and Physiology D-Genomics & Proteomics, v. 29, p. 95-105, MAR 2019. (10/05426-8, 13/09594-0, 12/25378-3)

AMARAL, DANILO T.; SILVA, JAQUELINE R.; VIVIANI, VADIM R.. RNA-Seq analysis of the bioluminescent and non-bioluminescent species of Elateridae (Coleoptera): Comparison to others photogenic and non-photogenic tissues of Elateroidea species. Comparative Biochemistry and Physiology D-Genomics & Proteomics, v. 29, p. 154-165, MAR 2019. (14/20176-9, 10/05426-8, 13/09594-0)

ROSA, SIMONE POLICENA; MARIANO, RAONE DOS REIS; VIVIANI, VADIM R.; COSTA, CLEIDE. Morphology of immature stages of Pyrearinus pumilus (Candeze, 1863) (Coleoptera: Elateridae: Pyrophorini): the click beetle responsible for the luminous canga caves in the state of Para, Brazil. Zootaxa, v. 4778, n. 3, p. 561-570, MAY 15 2020. (10/05426-8)

OLIVEIRA, GABRIELA; VIVIANI, VADIM R.. Comparison of the thermostability of recombinant luciferases from Brazilian bioluminescent beetles: Relationship with kinetics and bioluminescence colours. LUMINESCENCE, v. 33, n. 2, p. 282-288, MAR 2018. (15/05949-4, 10/05426-8)

CARVALHO, M. C.; TOMAZINI, A.; AMARAL, D. T.; MURAKAMI, M. T.; VIVIANI, V. R.. Luciferase isozymes from the Brazilian Aspisoma lineatum (Lampyridae) firefly: origin of efficient pH-sensitive lantern luciferases from fat body pH-insensitive ancestors. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 12, p. 1750-1764, DEC 1 2020. (10/05426-8, 16/07312-6)

AMARAL, DANILO T.; JOHNSON, CARL H.; VIVIANI, VADIM R.. RNA-Seq analysis of the blue light-emitting Orfelia fultoni (Diptera: Keroplatidae) suggest photoecological adaptations at the molecular level. Comparative Biochemistry and Physiology D-Genomics & Proteomics, v. 39, SEP 2021. (14/20176-9, 14/50583-5, 10/05426-8, 15/25051-2)

BEVILAQUA, V. R.; CARVALHO, MARIELE C.; PELENTIR, GABRIEL F.; TOMAZINI, ATILIO; MURAKAMI, MARIO; VIVIANI, VADIM R.. Influence of the C-terminal domain on the bioluminescence activity and color determination in green and red emitting beetle luciferases and luciferase-like enzyme. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 20, n. 1, JAN 2021. (10/05426-8)

VIVIANI, VADIM R.; SILVA, JAQUELINE R.; AMARAL, DANILO T.; BEVILAQUA, VANESSA R.; ABDALLA, FABIO C.; BRANCHINI, BRUCE R.; JOHNSON, CARL H.. A new brilliantly blue-emitting luciferin-luciferase system from Orfelia fultoni and Keroplatinae (Diptera). SCIENTIFIC REPORTS, v. 10, n. 1, JUN 15 2020. (10/05426-8, 14/50583-5)

CARVALHO, MARIELE C.; TOMAZINI, ATILIO; PRADO, ROGILENE A.; VIVIANI, VADIM R.. Selective inhibition of Zophobas morio (Coleoptera: Tenebrionidae) luciferase-like enzyme luminescence by diclofenac and potential suitability for light-off biosensing. LUMINESCENCE, v. 36, n. 2, NOV 2020. (16/07312-6, 14/18179-0, 10/05426-8)

VIVIANI, VADIM R.; BEVILAQUA, VANESSA R.; DE SOUZA, DANIEL R.; PELENTIR, GABRIEL F.; KAKIUCHI, MICHIO; HIRANO, TAKASHI. A Very Bright Far-Red Bioluminescence Emitting Combination Based on Engineered Railroad Worm Luciferase and 6 '-Amino-Analogs for Bioimaging Purposes. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 22, n. 1, p. 13-pg., 2021-01-01. (10/05426-8)

VIVIANI, V. R.; PELENTIR, G. F.; OLIVEIRA, G.; TOMAZINI, A.; BEVILAQUA, V. R.. Role of E270 in pH- and metal-sensitivities of firefly luciferases. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 11, p. 1548-1558, NOV 1 2020. (10/05426-8)

VIVIANI, VADIM R.; PELENTIR, GABRIEL F.; BEVILAQUA, VANESSA R.. Bioluminescence Color-Tuning Firefly Luciferases: Engineering and Prospects for Real-Time Intracellular pH Imaging and Heavy Metal Biosensing. BIOSENSORS-BASEL, v. 12, n. 6, p. 23-pg., 2022-06-01. (10/05426-8)

BEVILAQUA, V. R.; MATSUHASHI, T.; OLIVEIRA, G.; OLIVEIRA, P. S. L.; HIRANO, T.; VIVIANI, V. R.. Phrixotrix luciferase and 6 `-aminoluciferins reveal a larger luciferin phenolate binding site and provide novel far-red combinations for bioimaging purposes. SCIENTIFIC REPORTS, v. 9, JUN 21 2019. (10/05426-8)

FALASCHI, RAFAELA L.; AMARAL, DANILO T.; SANTOS, ISAIAS; DOMINGOS, ADAO H. R.; JOHNSON, GRANT A.; MARTINS, ANA G. S.; VIROOMAL, IMRAN B.; POMPEIA, SERGIO L.; MIRZA, JEREMY D.; OLIVEIRA, ANDERSON G.; et al. Neoceroplatus betaryiensis nov. sp. (Diptera: Keroplatidae) is the first record of a bioluminescent fungus-gnat in South America. SCIENTIFIC REPORTS, v. 9, AUG 5 2019. (10/05426-8, 17/22501-2, 13/16885-1)

BELLETI, ELISANGELA; BEVILAQUA, VANESSA R.; BRITO, ADRIANNE M. M.; MODESTO, DIEGO A.; LANFREDI, ALEXANDRE J. C.; VIVIANI, VADIM R.; NANTES-CARDOSO, ISELI L.. Synthesis of bioluminescent gold nanoparticle-luciferase hybrid systems for technological applications. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 20, n. 11, OCT 2021. (10/05426-8, 17/02317-2, 15/17688-0)

DE MELLO GABRIEL, GABRIELE VERONICA; YASUNO, RIE; MITANI, YASUO; OHMIYA, YOSHIHIRO; VIVIANI, VADIM RAVARA. Novel application of Macrolampis sp2 firefly luciferase for intracellular pH-biosensing in mammalian cells. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 18, n. 5, p. 1212-1217, MAY 1 2019. (10/05426-8, 14/04477-9, 16/15946-5, 11/23961-0, 15/22603-4)

OLIVEIRA, GABRIELA; VIVIANI, VADIM R.. Temperature effect on the bioluminescence spectra of firefly luciferases: potential applicability for ratiometric biosensing of temperature and pH. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 18, n. 11, p. 2682-2687, NOV 1 2019. (10/05426-8, 15/05949-4)

VIVIANI, VADIM R.; AMARAL, DANILO T.; BEVILAQUA, VANESSA R.; FALASCHI, RAFAELA L.. Orfelia-type luciferin and its associated storage protein in the non-luminescent cave worm Neoditomyia sp (Diptera: Keroplatidae) from the Atlantic rainforest: biological and evolutionary implications. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 17, n. 10, p. 1282-1288, OCT 1 2018. (10/05426-8)

VIVIANI, VADIM R.; GABRIEL, GABRIELE V. M.; BEVILAQUA, VANESSA R.; SIMOES, A. F.; HIRANO, T.; LOPES-DE-OLIVEIRA, P. S.. The proton and metal binding sites responsible for the pH-dependent green-red bioluminescence color tuning in firefly luciferases. SCIENTIFIC REPORTS, v. 8, DEC 4 2018. (10/05426-8, 13/09594-0)

VIVIANI, VADIM R.; SILVA, JAQUELINE RODRIGUES; HO, PAULO LEE. A Novel Brighter Bioluminescent Fusion Protein Based on ZZ Domain and Amydetes vivianii Firefly Luciferase for Immunoassays. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 9, OCT 18 2021. (10/05426-8)

MAZUMDER, ZEAUL H.; SHARMA, DEBDULAL; SENGUPTA, DEVASHISH; MUKHERJEE, AVINABA; BORUAH, JAYANTA SARMAH; BASU, SAMITA; SHUKLA, PRADEEP KUMAR; JHA, TARUN. Luciferase isozymes from the Brazilian Aspisoma lineatum (Lampyridae) firefly: origin of efficient pH-sensitive lantern luciferases from fat body pH-insensitive ancestors. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 12, p. 1776-1789, DEC 1 2020. (10/05426-8, 16/07312-6)

TOMAZINI, ATILIO; CARVALHO, MARIELE; MURAKAMI, MARIO T. T.; VIVIANI, VADIM R.. Effect of pH on the secondary structure and thermostability of beetle luciferases: structural origin of pH-insensitivity. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. N/A, p. 12-pg., 2023-01-21. (10/05426-8, 18/02538-1)

VIVIANI, VADIM R.; ROSA, SIMONE P.; PRADO, ROGILENE A.; PELENTIR, GABRIEL F.; DE SOUZA, DANIEL R.; REIS, RAONE M.; BECHARA, E. J. H.; COSTA, C.. Inventory and ecological aspects of bioluminescent beetles in the Cerrado ecosystem and its decline around Emas National Park (Brazil). Annals of the Entomological Society of America, v. N/A, p. 18-pg., 2023-10-04. (10/05426-8, 22/04800-0)

VIVIANI, VADIM R.; BEVILAQUA, VANESSA R.; DE SOUZA, DANIEL R.; PELENTIR, GABRIEL F.; KAKIUCHI, MICHIO; HIRANO, TAKASHI. A Very Bright Far-Red Bioluminescence Emitting Combination Based on Engineered Railroad Worm Luciferase and 6 `-Amino-Analogs for Bioimaging Purposes. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 22, n. 1, JAN 2021. (10/05426-8)

AMARAL, DANILO TRABUCO; MITANI, YASUO; BONATELLI, ISABEL APARECIDA SILVA; CERRI, RICARDO; OHMIYA, YOSHIHIRO; VIVIANI, VADIM RAVARA. Genome analysis of Phrixothrix hirtus (Phengodidae) railroad worm shows the expansion of odorant-binding gene families and positive selection on morphogenesis and sex determination genes. Gene, v. 850, p. 10-pg., 2023-01-20. (10/05426-8, 14/20176-9)

CARVALHO, M. C.; TOMAZINI, A.; AMARAL, D. T.; MURAKAMI, M. T.; VIVIANI, V. R.. Luciferase isozymes from the Brazilian Aspisoma lineatum (Lampyridae) firefly: origin of efficient pH-sensitive lantern luciferases from fat body pH-insensitive ancestors. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 12, p. 15-pg., 2020-12-01. (16/07312-6, 10/05426-8)

BEVILAQUA, V. R.; MATSUHASHI, T.; OLIVEIRA, G.; OLIVEIRA, P. S. L.; HIRANO, T.; VIVIANI, V. R.. Phrixotrix luciferase and 6 '-aminoluciferins reveal a larger luciferin phenolate binding site and provide novel far-red combinations for bioimaging purposes. SCIENTIFIC REPORTS, v. 9, p. 17-pg., 2019-06-21. (10/05426-8)

Filed patent(s) as a result of this research project