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

Author(s)

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

Affiliation

^[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

• Document type: Journal article
• Source: LUMINESCENCE; v. 33, n. 2, p. 282-288, MAR 2018.
• Web of Science Citations: 5

Abstract

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)

• FAPESP's process: 15/05949-4 - Development of far-red light emit thermostable luciferases of Phrixotrix hirtus for bioanalytical purposes: relation structure and function
• Grantee: Gabriela de Oliveira
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 10/05426-8 - Arthropod bioluminescence: biological diversity in Brazilian biomes, biochemical origin, structural/functional evolution of luciferases, molecular differentiation of lanterns, biotechnological, environmental and educational applications
• Grantee: Vadim Viviani
• Support Opportunities: Research Projects - Thematic Grants