Heterologous expression of luciferases in yeasts: potential applications in cell biosensors and bioluminescent immunoassays

• Grant number: 18/19430-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2018
• End date: October 31, 2021
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Ariele Cristina Moreira
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil
• Associated research grant: 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, AP.TEM

Abstract

Luciferases are the enzymes responsible for the production of bioluminescence in different organisms like bacteria, fungi, coelenterates, annelids, fish and insects. The beetle luciferases are among the most studied and used for biotechnological purposes. Many of these enzymes do not undergo post-translational modifications. However, some luciferases are glycosylated. Some insect luciferases cloned by our group have not yet been expressed in the active form in bacterial cells, among them are some luciferases of phengodids, and luciferases of bioluminescent dipterans. In order to express these luciferases in the active form, it is believed that eukaryotic cells, because they display post-translational modification machinery, are more suitable. Among heterologous expression systems of eukaryotes, the one of yeasts stands out for the simplicity, the lesser necessary infrastructure and cost. In addition, there is an interest in expressing luciferases in these cell types to generate eukaryotic cell biosensors for fermentation and biomass production processes. In this project it is then planned to subclone the cDNA encoding the above luciferases into yeast expression vectors, and express these enzymes in the active form for their characterization. It is also intended to test the viability of using yeasts transformed with these luciferases as bioluminescent cell biosensors.