Please use this identifier to cite or link to this item:
Full metadata record
DC FieldValueLanguage
dc.contributor.authordos Santos, Nathalia V.pt_PT
dc.contributor.authorMartins, Margaridapt_PT
dc.contributor.authorSantos-Ebinuma, Valéria C.pt_PT
dc.contributor.authorVentura, Sónia P. M.pt_PT
dc.contributor.authorCoutinho, João A. P.pt_PT
dc.contributor.authorValentini, Sandro R.pt_PT
dc.contributor.authorPereira, Jorge F. B.pt_PT
dc.description.abstractGreen fluorescent protein (GFP) has excellent properties as a biosensor and biomarker; however, its widespread use is limited by its purification costs. Alternative low-cost purification techniques can overcome this issue. The aim of this work was to evaluate aqueous biphasic systems (ABS) composed of cholinium chloride ([Ch]Cl) and different polymers as effective platforms to recover GFP from cell lysate of recombinant Escherichia coli BL21. All systems completely extracted GFP from cell lysate (>99%) into the polymeric- or [Ch]Cl-rich phases. In general, [Ch]Clbased ABS allowed a good purification capacity (GFP 80− 100% pure), with the best results (approximately 100% pure GFP) achieved with a polypropylene glycol (PPG)-400/[Ch]Cl ABS in a single-step extraction or in a two-step extraction (backextraction) by the integration of a polyethylene glycol (PEG)/sodium polyacrylate+[Ch]Cl ABS with a following stage using a PEG/[Ch]Cl-based ABS. Additionally, to demonstrate the potential of the PPG-400/[Ch]Cl ABS in downstream processing, solvent recycling and GFP polishing were carried out using ultrafiltration. Finally, the capacity of the PPG-400/[Ch]Cl ABS to extract other fluorescent proteins was also confirmed. The results clearly demonstrated that the PPG-400/[Ch]Cl ABS can be applied in downstream processing for the purification of proteins, not only enhancing purification yields but also providing simpler, quicker, cost-effective, and biocompatible processes.pt_PT
dc.publisherAmerican Chemical Societypt_PT
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FCTM%2F50011%2F2013/PTpt_PT
dc.subjectAqueous biphasic systemspt_PT
dc.subjectCholinium chloridept_PT
dc.subjectGreen fluorescent proteinpt_PT
dc.subjectIntegrated downstream processpt_PT
dc.titleAqueous biphasic systems composed of cholinium chloride and polymers as effective platforms for the purification of recombinant green fluorescent proteinpt_PT
degois.publication.titleACS Sustainable Chemistry and Engineeringpt_PT
Appears in Collections:CICECO - Artigos

Files in This Item:
File Description SizeFormat 
T.pdf2.76 MBAdobe PDFrestrictedAccess

Formato BibTex MendeleyEndnote Degois 

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.