Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros

Detalhes bibliográficos
Autor(a) principal: Souza, Daniel Rangel de
Data de Publicação: 2023
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/17527
Resumo: The origin and evolution of bioluminescence in Coleoptera from the Elateroidea superfamily depend on a better understanding of the metabolic origin of the luciferin substrate and the luciferase enzymes evolution. Several luciferases from the 4 bioluminescent beetles families (Lampyridae, Elateridae, Phengodidae and Rhagophthalmidae) have already been cloned and investigated by our and other research groups in recent decades. However, only three functional luciferases from the Phengodidae family have been cloned and investigated so far, the red light-emitting luciferase from Phrixotrix hirtus head and the green light-emitting luciferase from Phrixotrix vivianii. Thus, the origin and evolution of head and lateral lanterns luciferases, as well as the relationship between structure and bioluminescence spectra in these enzymes, are still incomplete, needing to study other luciferases from different lanterns and life stages of other species that emit intermediate colors of light. Within this theme, this thesis had two main goals: (1) Increase knowledge of the bioluminescence evolution in the Phengodidae family through cloning, expression and characterization new active luciferases cDNA from different lanterns and (2) understand the biosynthetic origin of beetle luciferin. For the first time, we associate larva and adult of a phengodid specie, the Stenophrixotrix pallens found on the UFSCar Sorocaba campus, through molecular analyzes of the 16S and 18S rRNA genes and the bioluminescence spectra. We cloned two new luciferases cDNA, one from the Brasilocerus sp. head and another from P. hirtus lateral lanterns, we expressed them in bacteria and characterized these luciferases. Luciferase from Brasilocerus sp. head lantern presented a peak of bioluminescent emission at 566 nm in pH=8, while the luciferase from the P. hirtus lateral lanterns shows an emission peak at 563 nm. The presence of a single luciferase in the head and lateral lanterns trascriptomes of Brasilocerus sp., the kinetic and spectral characteristics of luciferass from Stenophrixotrix pallens larvae, suggest for the first time that some species of Mastinocerini presente only one luciferase isoform in different lanterns and life stages, unlike Phrixotrix spp. The R215T mutation and the asparagine 351 deletion in the P.hirtus lateral lanterns luciferase had a bathochromic efect of ~30 nm, confirming the importance of arginine 215 and loop 351-360 in bioluminescence spectra determination. Finally, to better understand the biosynthetic origin of luciferin in coleopterans, we developed bioluminescent bacteria transformed with Amydetes vivianii luciferase to evaluate the in vivo synthesis of luciferin in the presence of cystein and quinine precursors. These studies showed that cysteine has a detoxifying impact on p-benzoquinone concomitantly with luciferin formation, suggesting that coleopteran luciferin may have arisen as a by-product of quinone detoxification.
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spelling Souza, Daniel Rangel deViviani, Vadim Ravarahttp://lattes.cnpq.br/3942043373510668http://lattes.cnpq.br/0245966248392912fa36fa1f-62a5-434f-abca-6d1b632098572023-03-23T16:42:08Z2023-03-23T16:42:08Z2023-03-09SOUZA, Daniel Rangel de. Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros. 2023. Tese (Doutorado em Biotecnologia e Monitoramento Ambiental) – Universidade Federal de São Carlos, Sorocaba, 2023. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17527.https://repositorio.ufscar.br/handle/ufscar/17527The origin and evolution of bioluminescence in Coleoptera from the Elateroidea superfamily depend on a better understanding of the metabolic origin of the luciferin substrate and the luciferase enzymes evolution. Several luciferases from the 4 bioluminescent beetles families (Lampyridae, Elateridae, Phengodidae and Rhagophthalmidae) have already been cloned and investigated by our and other research groups in recent decades. However, only three functional luciferases from the Phengodidae family have been cloned and investigated so far, the red light-emitting luciferase from Phrixotrix hirtus head and the green light-emitting luciferase from Phrixotrix vivianii. Thus, the origin and evolution of head and lateral lanterns luciferases, as well as the relationship between structure and bioluminescence spectra in these enzymes, are still incomplete, needing to study other luciferases from different lanterns and life stages of other species that emit intermediate colors of light. Within this theme, this thesis had two main goals: (1) Increase knowledge of the bioluminescence evolution in the Phengodidae family through cloning, expression and characterization new active luciferases cDNA from different lanterns and (2) understand the biosynthetic origin of beetle luciferin. For the first time, we associate larva and adult of a phengodid specie, the Stenophrixotrix pallens found on the UFSCar Sorocaba campus, through molecular analyzes of the 16S and 18S rRNA genes and the bioluminescence spectra. We cloned two new luciferases cDNA, one from the Brasilocerus sp. head and another from P. hirtus lateral lanterns, we expressed them in bacteria and characterized these luciferases. Luciferase from Brasilocerus sp. head lantern presented a peak of bioluminescent emission at 566 nm in pH=8, while the luciferase from the P. hirtus lateral lanterns shows an emission peak at 563 nm. The presence of a single luciferase in the head and lateral lanterns trascriptomes of Brasilocerus sp., the kinetic and spectral characteristics of luciferass from Stenophrixotrix pallens larvae, suggest for the first time that some species of Mastinocerini presente only one luciferase isoform in different lanterns and life stages, unlike Phrixotrix spp. The R215T mutation and the asparagine 351 deletion in the P.hirtus lateral lanterns luciferase had a bathochromic efect of ~30 nm, confirming the importance of arginine 215 and loop 351-360 in bioluminescence spectra determination. Finally, to better understand the biosynthetic origin of luciferin in coleopterans, we developed bioluminescent bacteria transformed with Amydetes vivianii luciferase to evaluate the in vivo synthesis of luciferin in the presence of cystein and quinine precursors. These studies showed that cysteine has a detoxifying impact on p-benzoquinone concomitantly with luciferin formation, suggesting that coleopteran luciferin may have arisen as a by-product of quinone detoxification.A compreensão da origem e evolução da bioluminescência em coleópteros da superfamília Elateroidea, depende de um melhor entendimento da origem metabólica do substrato luciferina e da evolução das enzimas luciferases. Diversas luciferases de espécies das 4 famílias de coleópteros bioluminescentes (Lampyridae, Elateridae, Phengodidae e Rhagophthalmidae) já foram clonadas e investigadas por nosso e outros grupos de pesquisa nas últimas décadas. No entanto, apenas três luciferases funcionais da família Phengodidae foram clonadas e investigadas até agora, a luciferase emissora de luz vermelha da cabeça de Phrixotrix hirtus e a luciferase emissora de luz verde de Phrixotrix vivianii. Assim, a origem e evolução das luciferases das lanternas da cabeça e do corpo, bem como a relação entre estrutura e espectros de bioluminescência nestas enzimas, ainda estão incompletas, havendo a necessidade do estudo de outras luciferases oriundas das diferentes lanternas e estágios de vida de outras espécies que emitem cores intermediárias de luz. Dentro dessa temática, esta tese teve dois objetivos principais: (1) Aumentar o conhecimento da evolução da biologia e bioluminescência da família Phengodidae através da clonagem, expressão e caracterização dos cDNAs de novas luciferases ativas oriundas de diferentes lanternas e (2) entender a origem biossintética da luciferina de besouros. Pela primeira vez, conseguimos associar larva e adulto de uma espécie de fengodídeo, o Stenophrixotrix pallens encontrado no campus da UFSCar Sorocaba, através de análises moleculares dos genes 16S e 18S rRNA e dos espectros de bioluminescência. Clonamos os cDNAs de duas novas luciferases, uma oriunda da cabeça de Brasilocerus sp. e outra das lanternas laterais de P. hirtus, expressamos os mesmos em bactérias e caracterizamos estas luciferases. A luciferase da lanterna cefálica de Brasilocerus sp. apresentou um pico de emissão bioluminescente em 566 nm em pH=8, enquanto que a luciferase das lanternas laterais de P. hirtus apresenta um pico de emissão em 563 nm. A presença de uma única luciferase nos transcriptomas das lanternas laterais e da cabeça em Brasilocerus, as características espectrais das luciferases de larvas de Stenophrixotrix pallens, sugerem pela primeira vez que algumas espécies de Mastinocerini apresentam apenas uma isoforma de luciferase em diferentes lanternas e estágios de vida, diferentemente de Phrixotrix spp. As mutações R215T e a deleção da asparagina 351 na luciferase das lanternas laterais de P. hirtus, tiveram um efeito batocrômico de ~30 nm, confirmando a importância da arginina 215 e do loop 351-360 na determinação dos espectros de bioluminescência. Finalmente, para melhor entendermos a origem biossintética da luciferina em coleópteros, desenvolvemos bactérias bioluminescentes transformadas com a luciferase de Amydetes vivianii para avaliar a síntese in vivo de luciferina na presença dos precursores cisteína e quinonas. Estes estudos mostraram que a cisteína tem efeito detoxificante de p-benzoquinona concomitantemente com a formação de luciferina, sugerindo que a luciferina de coleópteros possa ter surgido como subproduto da detoxificação de quinonas.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)2018/17855-2porUniversidade Federal de São CarlosCâmpus SorocabaPrograma de Pós-Graduação em Biotecnologia e Monitoramento Ambiental - PPGBMA-SoUFSCarAttribution 3.0 Brazilhttp://creativecommons.org/licenses/by/3.0/br/info:eu-repo/semantics/openAccessBioluminescênciaFengodídeosLuciferasesLuciferinaBioluminescencePhengodidaeLuciferinCIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAREstrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópterosStructure, function, and evolution of luciferases from Mastinocerini (Coleoptera: Phengodidae) and luciferin biosynthetic origin in coleopterainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis600600fe330127-59af-4e10-b5a5-580f91a07359reponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTese_DanielRangeldeSouza.pdfTese_DanielRangeldeSouza.pdfTese de Doutoradoapplication/pdf3214911https://repositorio.ufscar.br/bitstream/ufscar/17527/1/Tese_DanielRangeldeSouza.pdf6d25c08a9ecae202acfb722fb0a467dcMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8913https://repositorio.ufscar.br/bitstream/ufscar/17527/2/license_rdf3185b4de2190c2d366d1d324db01f8b8MD52TEXTTese_DanielRangeldeSouza.pdf.txtTese_DanielRangeldeSouza.pdf.txtExtracted texttext/plain178720https://repositorio.ufscar.br/bitstream/ufscar/17527/3/Tese_DanielRangeldeSouza.pdf.txte455efab2d70e441b383fa14048b91e0MD53THUMBNAILTese_DanielRangeldeSouza.pdf.jpgTese_DanielRangeldeSouza.pdf.jpgIM Thumbnailimage/jpeg6645https://repositorio.ufscar.br/bitstream/ufscar/17527/4/Tese_DanielRangeldeSouza.pdf.jpgd1a4b90fe538879cbaeea11e586b8d11MD54ufscar/175272023-09-18 18:32:34.719oai:repositorio.ufscar.br:ufscar/17527Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:32:34Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
dc.title.alternative.eng.fl_str_mv Structure, function, and evolution of luciferases from Mastinocerini (Coleoptera: Phengodidae) and luciferin biosynthetic origin in coleoptera
title Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
spellingShingle Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
Souza, Daniel Rangel de
Bioluminescência
Fengodídeos
Luciferases
Luciferina
Bioluminescence
Phengodidae
Luciferin
CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
title_short Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
title_full Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
title_fullStr Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
title_full_unstemmed Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
title_sort Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros
author Souza, Daniel Rangel de
author_facet Souza, Daniel Rangel de
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/0245966248392912
dc.contributor.author.fl_str_mv Souza, Daniel Rangel de
dc.contributor.advisor1.fl_str_mv Viviani, Vadim Ravara
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/3942043373510668
dc.contributor.authorID.fl_str_mv fa36fa1f-62a5-434f-abca-6d1b63209857
contributor_str_mv Viviani, Vadim Ravara
dc.subject.por.fl_str_mv Bioluminescência
Fengodídeos
Luciferases
Luciferina
topic Bioluminescência
Fengodídeos
Luciferases
Luciferina
Bioluminescence
Phengodidae
Luciferin
CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
dc.subject.eng.fl_str_mv Bioluminescence
Phengodidae
Luciferin
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
description The origin and evolution of bioluminescence in Coleoptera from the Elateroidea superfamily depend on a better understanding of the metabolic origin of the luciferin substrate and the luciferase enzymes evolution. Several luciferases from the 4 bioluminescent beetles families (Lampyridae, Elateridae, Phengodidae and Rhagophthalmidae) have already been cloned and investigated by our and other research groups in recent decades. However, only three functional luciferases from the Phengodidae family have been cloned and investigated so far, the red light-emitting luciferase from Phrixotrix hirtus head and the green light-emitting luciferase from Phrixotrix vivianii. Thus, the origin and evolution of head and lateral lanterns luciferases, as well as the relationship between structure and bioluminescence spectra in these enzymes, are still incomplete, needing to study other luciferases from different lanterns and life stages of other species that emit intermediate colors of light. Within this theme, this thesis had two main goals: (1) Increase knowledge of the bioluminescence evolution in the Phengodidae family through cloning, expression and characterization new active luciferases cDNA from different lanterns and (2) understand the biosynthetic origin of beetle luciferin. For the first time, we associate larva and adult of a phengodid specie, the Stenophrixotrix pallens found on the UFSCar Sorocaba campus, through molecular analyzes of the 16S and 18S rRNA genes and the bioluminescence spectra. We cloned two new luciferases cDNA, one from the Brasilocerus sp. head and another from P. hirtus lateral lanterns, we expressed them in bacteria and characterized these luciferases. Luciferase from Brasilocerus sp. head lantern presented a peak of bioluminescent emission at 566 nm in pH=8, while the luciferase from the P. hirtus lateral lanterns shows an emission peak at 563 nm. The presence of a single luciferase in the head and lateral lanterns trascriptomes of Brasilocerus sp., the kinetic and spectral characteristics of luciferass from Stenophrixotrix pallens larvae, suggest for the first time that some species of Mastinocerini presente only one luciferase isoform in different lanterns and life stages, unlike Phrixotrix spp. The R215T mutation and the asparagine 351 deletion in the P.hirtus lateral lanterns luciferase had a bathochromic efect of ~30 nm, confirming the importance of arginine 215 and loop 351-360 in bioluminescence spectra determination. Finally, to better understand the biosynthetic origin of luciferin in coleopterans, we developed bioluminescent bacteria transformed with Amydetes vivianii luciferase to evaluate the in vivo synthesis of luciferin in the presence of cystein and quinine precursors. These studies showed that cysteine has a detoxifying impact on p-benzoquinone concomitantly with luciferin formation, suggesting that coleopteran luciferin may have arisen as a by-product of quinone detoxification.
publishDate 2023
dc.date.accessioned.fl_str_mv 2023-03-23T16:42:08Z
dc.date.available.fl_str_mv 2023-03-23T16:42:08Z
dc.date.issued.fl_str_mv 2023-03-09
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dc.identifier.citation.fl_str_mv SOUZA, Daniel Rangel de. Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros. 2023. Tese (Doutorado em Biotecnologia e Monitoramento Ambiental) – Universidade Federal de São Carlos, Sorocaba, 2023. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17527.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/17527
identifier_str_mv SOUZA, Daniel Rangel de. Estrutura, função e evolução das luciferases de Mastinocerini (Coleoptera: Phengodidae) e origem biossintética da luciferina em coleópteros. 2023. Tese (Doutorado em Biotecnologia e Monitoramento Ambiental) – Universidade Federal de São Carlos, Sorocaba, 2023. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17527.
url https://repositorio.ufscar.br/handle/ufscar/17527
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dc.rights.driver.fl_str_mv Attribution 3.0 Brazil
http://creativecommons.org/licenses/by/3.0/br/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution 3.0 Brazil
http://creativecommons.org/licenses/by/3.0/br/
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dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus Sorocaba
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Biotecnologia e Monitoramento Ambiental - PPGBMA-So
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publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus Sorocaba
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