Structure and biological function of luciferase-like enzime of Arachnocampa luminosa lanterns (Diptera: Keroplatidae) and luminescent activity development by genetic engineering

• Grant number: 16/07312-6
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: July 01, 2016
• End date: July 31, 2020
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Mariele Cristina de Carvalho
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil

Abstract

The bioluminescence in different organisms arises from the exothermic reaction in which enzymes known as luciferases catalyze the oxidation of organic substrates known as luciferins, resulting in electronically excited products and production of light with high efficiency. Several bioluminescente systems have been investigated and their biochemistry is well known. In Diptera, two distinct bioluminescent systems are found, Arachnocampa and Orfelia. In Arachnocampa, the system involves an unknown luciferase and luciferin and ATP. Previously, the supervisor of this project identified through proteomic analysis a luciferase-like enzyme similar to beetle luciferases in purified lanterns extracts of Arachnocampa luminosa, and then cloned the cDNA for this enzyme. However, this enzyme showed no Luminescent activity, meaning the function of this enzyme in the lanterns and it's relationship in the bioluminescence remains unclear. Considering that some luciferase-like enzymes exhibit luminescence activity with fireflies’ luciferin, and the potential shown to increase the luminescent activity in some of these enzymes, we plan to investigate the natural function of this luciferase-like enzyme and develop Luminescent activity by Genetic Engineering. Thus, this project aims to investigate the function of this luciferase-like enzyme from Arachnocampa lanterns using: (1) luminescence tests with fireflies and Arachnocampa luciferin; (2) tioesterification assays with different classes of carboxylic acids aimed at identifying the preferred type of substrate; and (3) to develop luminescence activity with firefly luciferin by Genetic Engineering using site-direct and Random mutagenesis. We will also investigate other luciferase-like enzymes found in Arachnocampa lanterns. (AU)

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MAZUMDER, ZEAUL H.; SHARMA, DEBDULAL; SENGUPTA, DEVASHISH; MUKHERJEE, AVINABA; BORUAH, JAYANTA SARMAH; BASU, SAMITA; SHUKLA, PRADEEP KUMAR; JHA, TARUN. Luciferase isozymes from the Brazilian Aspisoma lineatum (Lampyridae) firefly: origin of efficient pH-sensitive lantern luciferases from fat body pH-insensitive ancestors. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 12, p. 1776-1789, DEC 1 2020. (10/05426-8, 16/07312-6)

CARVALHO, M. C.; TOMAZINI, A.; AMARAL, D. T.; MURAKAMI, M. T.; VIVIANI, V. R.. Luciferase isozymes from the Brazilian Aspisoma lineatum (Lampyridae) firefly: origin of efficient pH-sensitive lantern luciferases from fat body pH-insensitive ancestors. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 12, p. 15-pg., 2020-12-01. (16/07312-6, 10/05426-8)

CARVALHO, M. C.; TOMAZINI, A.; AMARAL, D. T.; MURAKAMI, M. T.; VIVIANI, V. R.. Luciferase isozymes from the Brazilian Aspisoma lineatum (Lampyridae) firefly: origin of efficient pH-sensitive lantern luciferases from fat body pH-insensitive ancestors. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 19, n. 12, p. 1750-1764, DEC 1 2020. (10/05426-8, 16/07312-6)

CARVALHO, MARIELE C.; TOMAZINI, ATILIO; PRADO, ROGILENE A.; VIVIANI, VADIM R.. Selective inhibition of Zophobas morio (Coleoptera: Tenebrionidae) luciferase-like enzyme luminescence by diclofenac and potential suitability for light-off biosensing. LUMINESCENCE, v. 36, n. 2, NOV 2020. (16/07312-6, 14/18179-0, 10/05426-8)