R-phycoerythrin extraction and purification from fresh Gracilaria sp. using thermo-responsive systems

abstract

R-phycoerythrin is a high added-value protein found in red macroalgae with several interesting properties. Despite the promising results found when R-phycoerythrin is used as an optically active center in luminescent solar concentrators (LSCs), it still has some problems that can be attributed to the low stability of the R-phycoerythrin in the presence of the specific contaminant proteins found in the crude extract. The development of downstream strategies able to reduce the use of environmentally hazardous solvents, while improving the purification without compromising the R-phycoerythrin structural integrity is still the biggest challenge to overcome. Aqueous micellar two-phase systems (AMTPS) appear as an appealing fractionation approach since they allow the processing of systems with larger water contents, while displaying great selectivity and biocompatibility with several biomolecules. Moroever, AMTPS that mix surfactants and surface-active ionic liquids are shown to significantly enhance protein purification. In this work, mixed AMTPS were applied to the R-phycoerythrin purification from red macroalgae. After the process optimization, this work proposes the application of two consecutive steps of purification as the final process to isolate R-phycoerythrin from the remaining proteins composing the crude extract, while maintaining the structural integrity of R-phycoerythrin, as requested to be used in the LSC. Besides a good performance, the two-step approach developed was also shown to have a lower environmental impact with a carbon footprint decrease of 16%, when compared with the conventional AMTPS.

keywords

GRATELOUPIA-TURUTURU; 2-PHASE SYSTEMS; IONIC LIQUIDS; CYTOCHROME-C; PROTEINS; SURFACTANTS; SEPARATION

subject category

Chemistry; Science & Technology - Other Topics

authors

Vicente, FA; Cardoso, IS; Martins, M; Goncalves, CVM; Dias, ACRV; Domingues, P; Coutinho, JAP; Ventura, SPM

Groups

G4 - Renewable Materials and Circular Economy
G5 - Biomimetic, Biological and Living Materials

Projects

CICECO - Aveiro Institute of Materials (UID/CTM/50011/2019)

SusPhotoSolutions - Soluções Fotovoltaicas Sustentáveis (SUSPHOTOSOLUTIONS)

Projeto de Investigação Exploratória: Sónia Patricia Marques Ventura (IF/00402/2015)

acknowledgements

This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, FCT Ref. UID/CTM/50011/2019, financed by national funds through FCT/MCTES, CESAM (UID/AMB/50017 -POCI-01-0145-FEDER-007638), and QOPNA research unit (FCT UID/QUI/00062/2013) financed by national funds through FCT/MEC and when appropriate co-financed by the FEDER under the PT2020 Partnership Agreement. It was also supported by the Integrated Programme of SR&TD SusPhotoSolutions -Solucoes Fotovoltaicas Sustentaveis (reference CENTRO-01-0145-FEDER-000005), co-funded by the Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. The authors are grateful for the national fund through the Portuguese Foundation for Science and Technology (FCT) for the doctoral grants SFRH/BD/101683/2014 of F.A. Vicente and SFRH/BD/122220/2016 of M. Martins. S. P. M. Ventura and A. C. R. V. Dias acknowledge FCT for the contracts IF/00402/2015 and IF/00587/2013, respectively. Thanks are also due to the Fundacao para a Ciencia e a Tecnologia (FCT, Portugal), for funding RNEM (REDE/1504/REM/2005) the Portuguese Mass Spectrometry Network. The authors thank ALGAplus company for the macroalgae samples.