Extraction of recombinant proteins from Escherichia coli by cell disruption with aqueous solutions of surface-active compounds


BACKGROUNDGreen fluorescent protein (GFP) is used extensively as a biomarker due to its unique spectral and fluorescence characteristics. GFP is usually obtained from recombinant strains of Escherichia coli (E. coli) producing the protein intracellularly. However, the common methods of extraction are cumbersome leading to an increase in the downstream process costs and complexity, sometimes leading to a higher risk of biomolecule degradation. RESULTSThis work proposes a new method to extract recombinant intracellular GFP from E. coli BL21 by using aqueous solutions of surface-active compounds. CONCLUSIONBy comparing the fluorescence intensity of the extracted GFP, it was concluded that some of these compounds, namely ionic liquids with an alkyl chain of 10 or more carbons (best solvent being tributyl-1-tetradecylphosphonium, [P-4,P-4,P-4,P-14]Cl) are more effective than an ultrasonic-assisted extraction, even at low concentrations, being able to extract the whole GFP content from the cells. (c) 2018 Society of Chemical Industry




Biotechnology & Applied Microbiology; Chemistry; Engineering


Martins, M; Ooi, CW; Neves, MC; Pereira, JFB; Coutinho, JAP; Ventura, SPM

nossos autores


This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, POCI-01-0145-FEDER-007679 (FCT Ref. UID/CTM/50011/2013), financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. The authors are also grateful to FCT for financial support via the Project "Optimization and Scale-up of Novel Ionic-Liquid-based Purification Processes for Recombinant Green Fluorescent Protein produced by Escherichia coli', process number FAPESP/19793/2014. This work was supported by the Integrated Programme of SR&TD 'SusPhotoSolutions - Solucoes Fotovoltaicas Sustentaveis' (reference CENTRO-01-0145-FEDER-000005), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. The authors also thank FCT for the post-doctoral grant SFRH/BPD/110423/2015 to MC Neves and doctoral grant SFRH/BD/122220/2016 to M Martins. SPM Ventura acknowledges the IF contract IF/00402/2015. JFB Pereira acknowledges financial support from FAPESP through the project 2014/16424-7. CW Ooi acknowledges funding support from the Ministry of Science, Technology and Innovation, Malaysia (e-Science funding: 02-02-10-SF0290).

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