Development of liquid-liquid purification process and incorporation in nanostructured polymeric matrix of recombinant green fluorescent protein produced by Escherichia coli

• Grant number: 16/07529-5
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: August 01, 2016
• End date: November 30, 2020
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Jorge Pereira
• Grantee: Nathalia Vieira Porphirio Veríssimo
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated scholarship(s): 18/06576-5 - Molecular interactions and stability of Green Fluorescent Protein (GFP) in ionic liquids and encapsulation of GFP on reverse micelles, BE.EP.DD

Abstract

The Green Fluorescent Protein (GFP) has excellent properties to act as a biosensor and biomarker, but its high cost and difficulties in application limits its use. The study of new methods of purification and protection/encapsulation of GFP can solve these limitations. With this in mind, the project aims to develop new methods for the purification of GFP and adapt them to industrial scale. For this purpose, we will evaluate Ionic-Liquid-based Aqueous Biphasic Systems (IL-based ABS) composed of a) polymers, salts and ionic liquids as adjuvants and b) polymers and ionic liquids, for the purification of GFP. The work also aims to incorporate the purified GFP in a nanostructured polymer matrix and test the stability and biocompatibility of the resulting nanomaterial. The main objective of the project is to improve and expand the application of GFP in the areas of Biotechnology, Pharmacy and Medicine. (AU)

Scientific publications

(9)

(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; SANTOS-EBINUMA, VALERIA DE CARVALHO; PESSOA JUNIOR, ADALBERTO; BRANDAO PEREIRA, JORGE FERNANDO. Liquid-liquid extraction of biopharmaceuticals from fermented broth: trends and future prospects. JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, v. 93, n. 7, SI, p. 1845-1863, JUL 2018. (14/16424-7, 14/01580-3, 14/19793-3, 13/08617-7, 16/07529-5)

TORRES-ACOSTA, MARIO A.; DOS SANTOS, V, NATHALIA; VENTURA, SONIA P. M.; COUTINHO, JOAO A. P.; RITO-PALOMARES, MARCO; PEREIRA, JORGE F. B.. Economic analysis of the production and recovery of green fluorescent protein using ATPS-based bioprocesses. Separation and Purification Technology, v. 254, JAN 1 2021. (14/16424-7, 14/19793-3, 16/07529-5)

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)

LOPES, CAMILA; DOS SANTOS, NATHALIA VIEIRA; DUPONT, JANA; PEDROLLI, DANIELLE BISCARO; VALENTINI, SANDRO ROBERTO; SANTOS-EBINUMA, VALERIA DE CARVALHO; BRANDAO PEREIRA, JORGE FERNANDO. Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration. Biotechnology and Applied Biochemistry, v. 66, n. 4, p. 527-536, JUL 2019. (14/16424-7, 18/50009-8, 14/19793-3, 16/07529-5)

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)

SOARES, BRUNA P.; SANTOS, JOAO H. P. M.; MARTINS, MARGARIDA; ALMEIDA, MAFALDA R.; SANTOS, V, NATHALIA; FREIRE, MARA G.; SANTOS-EBINUMA, VALERIA C.; COUTINHO, JOAO A. P.; PEREIRA, JORGE F. B.; VENTURA, SONIA P. M.. Purification of green fluorescent protein using fast centrifugal partition chromatography. Separation and Purification Technology, v. 257, FEB 15 2021. (14/16424-7, 18/25994-2, 16/07529-5)

DOS SANTOS, NATHALIA V.; MARTINS, MARGARIDA; SANTOS-EBINUMA, VALERIA C.; VENTURA, SONIA P. M.; COUTINHO, JOAO A. P.; VALENTINI, SANDRO R.; PEREIRA, JORGE F. B.. Aqueous Biphasic Systems Composed of Cholinium Chloride and Polymers as Effective Platforms for the Purification of Recombinant Green Fluorescent Protein. ACS SUSTAINABLE CHEMISTRY & ENGINEERING, v. 6, n. 7, p. 9383-9393, JUL 2018. (14/16424-7, 14/19793-3, 16/07529-5)

DOS SANTOS, NATHALIA VIEIRA; SANTOS-EBINUMA, VALERIA DE CARVALHO; PESSOA JUNIOR, ADALBERTO; BRANDAO PEREIRA, JORGE FERNANDO. Liquid-liquid extraction of biopharmaceuticals from fermented broth: trends and future prospects. JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, v. 93, n. 7, p. 19-pg., 2018-07-01. (14/16424-7, 14/01580-3, 16/07529-5, 13/08617-7, 14/19793-3)