Selection of thermostable mutants of Phrixotrix hirtus luciferase for bioanalytical purposes

• Grant number: 13/18449-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2013
• End date: October 31, 2014
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Gabriela de Oliveira
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil

Abstract

Luciferases are enzymes that catalyze the oxidization of molecules known as luciferins resulting in the production of bioluminescence. Among luciferases of different organisms, those found in fireflies and other beetles have been the focus of many studies and are currently applied for many biotechnological purposes, including bioanalysis of ATP, reporter gene assays and cellular bioimaging. The luciferases from beetles arequite sensitivity to high temperatures therefore it is necessary toimprove their thermos ability in order to apply them for bio imaging at high-temperature tissues of mammalians. The luciferase of Phrixotrixrailroadworm is especially interesting for hemoglobin rich tissues and pigmented samples, because is the only natural red emitting luciferase to be cloned. However, it is unstable at higher temperatures of mammalian tissues. The aim of this study is to engineer, select and characterize thermostable mutants of the unique red emitting Phrixotrix railroad worm luciferase through site-directed mutations.