Download Surface-active ionic liquids applied on the recovery of green

N°1206 / PC
TOPIC(s) : Alternative solvents / Biotechnologies
Surface-active ionic liquids applied on the recovery of green fluorescent protein
from Escherichia coli cells
AUTHORS
Sónia VENTURA / UNIVERSITY OF AVEIRO, CAMPUS UNIIVERSITÁRIO DE SANTIAGO, AVEIRO
PURPOSE OF THE ABSTRACT
The green fluorescent protein (GFP) is recognized as one of the most promising biomarkers, due to its unique
spectral and fluorescence characteristics. As the recombinant GFP is usually expressed intracellularly, for
example, by recombinant strains of Escherichia coli [1], a preliminary step of cell disruption is mandatory. The
conventional methods of cell disruption include the mechanical methods (e.g., multiple cycles of freezing/thawing
[2] and/or ultrasonic homogenization [3]) or the non-mechanical methods (such as the enzymatic treatments [4]).
However, the mechanical methods of cell disruption exhibit several drawbacks, namely the long processing time,
high energy consumption, high costs, and potential denaturing effects on the structural integrity of proteins. In this
sense, surface-active compounds can be used as membrane permeabilizing agents owing to the mild and simple
operation conditions. In this work, conventional surfactants and tensioactive ionic liquids were tested in term of
their potential as cell disruptor agents. Aqueous solutions of several surface-active compounds were used to
solubilize the lipidic membranes of the bacterial cell, leading to the GFP release into the extracellular medium.
The promising results were obtained, as the GFP was completely released from Escherichia coli, being the
fluorescence intensity of the extracts even improved in some cases.
Acknowledgments
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 by the financial support considering the Project ?Optimization and Scale-up of Novel
Ionic-Liquid-based Purification Processes for Recombinant Green Fluorescent Protein produced by Escherichia
coli?, process number FCT/FAPESP 2014/19793-3. S.P.M. Ventura thanks FCT for the financial support through
the contract IF/00402/2015. J.F.B. Pereira acknowledges financial support from FAPESP through the project
2014/19793-3. Ooi C.W. is thankful for the funding support from Ministry of Science, Technology and Innovation,
Malaysia (e-Science funding: 02-02-10-SF0290).
FIGURES
FIGURE 1
FIGURE 2
KEYWORDS
Surface-active ionic liquids | Extraction | Green fluorescent protein | Escherichia coli
BIBLIOGRAPHY
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