Molecular interactions and stability of Green Fluorescent Protein (GFP) in ionic liquids and encapsulation of GFP on reverse micelles

• Grant number: 18/06576-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: September 01, 2018
• End date: August 31, 2019
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Jorge Pereira
• Grantee: Nathalia Vieira Porphirio Veríssimo
• Supervisor: Tamar Greaves
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Institution abroad: RMIT University, Melbourne, Australia
• Associated to the scholarship: 16/07529-5 - Development of liquid-liquid purification process and incorporation in nanostructured polymeric matrix of recombinant green fluorescent protein produced by Escherichia coli, BP.DD

Abstract

Advances in biotechnology allowed the discovery of many bioproducts with medical applications, but the instability of some of these molecules in biological systems makes their application very challenging. Finding solvents or formulations that stabilize these biomolecules can be a solution for these difficulties, but greater understanding of the solvent-biomolecule interactions is needed. The goal of this project is to study the interactions that occur at a molecular level among aqueous solutions of different solvents (in particular, ionic liquids - ILs) with biomolecules. We have selected the Green Fluorescent Protein (GFP) which is an important biopharmaceutical molecule that is used as a biosensor and biomarker. This study will lead to greater comprehension of the interaction mechanism between proteins and ILs. Depending of the results of the stability studies, new ILs with more biocompatible properties can be rationally designed. Additionally, this work also aims to develop and characterize formulations for the encapsulation of GFP, specifically, reverse micelles. Both the stability and encapsulation studies have the major goal of improving and broadening the applications of GFP in the medical and industrial fields. The stability of GFP will be assessed in different solvents at various concentrations and stress conditions, with evaluation of its fluorescent intensity associated with in-depth studies of its secondary and tertiary structure using spectroscopy and light scattering techniques. The reverse micelles will be characterized using light scattering analysis and small angle scattering techniques, and the encapsulation efficiency and stability of the encapsulated GFP will also be evaluated. (AU)

Scientific publications

(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)

DOS SANTOS, NATHALIA VIEIRA; SAPONI, CAROLINA FALASCHI; GREAVES, TAMAR LOUISE; BRANDAO PEREIRA, JORGE FERNANDO. Revealing a new fluorescence peak of the enhanced green fluorescent protein using three-dimensional fluorescence spectroscopy. RSC ADVANCES, v. 9, n. 40, p. 22853-22858, AUG 1 2019. (14/16424-7, 18/50009-8, 18/06576-5, 18/01858-2, 14/19793-3, 16/07529-5)

DOS SANTOS, NATHALIA VIEIRA; SAPONI, CAROLINA FALASCHI; RYAN, TIMOTHY M.; PRIMO, FERNANDO L.; GREAVES, TAMAR L.; PEREIRA, JORGE F. B.. Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors. International Journal of Biological Macromolecules, v. 164, p. 3474-3484, DEC 1 2020. (18/20833-0, 14/16424-7, 14/19793-3, 18/06576-5, 16/07529-5, 18/50009-8, 18/01858-2)

V. VERISSIMO, NATHALIA; SAPONI, CAROLINA F.; RYAN, TIMOTHY M.; GREAVES, TAMAR L.; PEREIRA, JORGE F. B.. Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity. GREEN CHEMICAL ENGINEERING, v. 2, n. 4, p. 11-pg., 2021-12-01. (18/06576-5, 18/25511-1, 14/16424-7, 20/14144-8, 18/50009-8, 14/19793-3, 18/01858-2, 18/20833-0, 16/07529-5)