Comparison of the thermostability of recombinant luciferases from Brazilian bioluminescent beetles: Relationship with kinetics and bioluminescence colours

Autor(es)

Oliveira, Gabriela ^{[1, 2]} ; Viviani, Vadim R. ^{[1, 2]}

Afiliação do(s) autor(es)

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

^[2] Fed Univ Sao Carlos UFSCar, Dept Chem Phys & Math, Grad Program Biotechnol & Environm Monitoring, Lab Biochem & Biotechnol Bioluminescence, Campus Sorocaba, Sorocaba, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: LUMINESCENCE; v. 33, n. 2, p. 282-288, MAR 2018.
• Citações Web of Science: 5

Resumo

Firefly luciferases have been used extensively as bioanalytical reagents and their cDNAs as reporter genes for biosensors and bioimaging, but they are in general unstable at temperatures above 30 degrees C. In the past few years, efforts have been made to stabilize some firefly luciferases for better application as analytical reagents. Novel luciferases from different beetle families, displaying distinct bioluminescence colours and kinetics, may offer desirable alternatives to extend the range of applications. In the past years, our group has cloned the largest variety of luciferases from the three main families of bioluminescent beetles (Elateridae: P. termitilluminans, F. bruchi, P. angustus; Phengodidae: P. hirtus, P. vivianii; and Lampyridae: A. vivianii, C. distinctusand Macrolampis sp2) occurring in Brazilian biomes. We compared the thermostability of these recombinant luciferases and investigated their relationships with bioluminescence spectra and kinetics. The most thermostable luciferases were those of Pyrearinus termitilluminans larval click beetle (534nm), Amydetes vivianii firefly (539nm) and Phrixotrix vivianii railroad worm (546nm), which are the most blue-shifted examples in each family, confirming the trend that the most blue-shifted emitting luciferases are also the most thermostable. Comparatively, commercial P. pyralis firefly luciferase was less thermostable than P. termitilluminans click beetle and A. vivianii firefly luciferases. The higher thermostability in these luciferases could be related to higher degree of hydrophobic packing and disulfide bond content (for firefly luciferases). (AU)

• Processo FAPESP: 15/05949-4 - Desenvolvimento de luciferases emissoras de luz vermelha distante termoestáveis de Phrixotrix hirtus para finalidades bioanalíticas: relação estrutura e função
• Beneficiário: Gabriela de Oliveira
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• 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