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The temperature effect on electroextraction of proteins from bacterial cells

STSM by Saša Haberl Meglič, Early-Stage Researcher

Period of mission: from 23/03/2014 to 30/03/2014

Home institution: Laboratory of Biocybernetics, University of Ljubjana, Faculty of Electrical Engineering, Ljubljana, Slovenia

Host institution: University of Zaragoza, Faculty of Veterinary Medicine, Zaragoza, Spain

An increasing number of proteins are being used in medicine, food industry and biotechnology and the need to develop methods for extraction on a large production scale is becoming increasingly important. Different chemical or physical methods are currently used for extraction of proteins from bacterial cells. But on a large scale, a general handicap of all extraction processes is that concentration of contaminants is high, necessitating downstream processing. Extraction of bacterial proteins by means of electroporation has opened a new field for efficient and cost-effective protein production on a large scale and many parameters (e.g. pulse parameters, temperature, etc.) can influence the efficiency the method. Therefore, the main purpose of this STSM which was performed under supervision of Dr. Javier Raso was to test the influence of temperature during exposure of cells to electric pulses (10, 25 and 40°C) and post-pulse incubation temperature (25 and 37°C) and time (from 1 h to 24 h) on extraction of proteins by means of electroporation from bacterial cells. Also the influence of electric pulse parameters (electric field strength and pulse number) on bacterial membrane permeabilization, bacterial inactivation and the concentration of extracted proteins were observed. The model organism for our investigation was E. coli bacteria, strain K12 Top10 and our results showed that with increasing electric field strength, treatment time or treatment temperature also membrane permeabilization increases. Also increased electric field strength or treatment time increases bacteria inactivation. The most tremendous effect on the concentration of extracted proteins by means of electroporation was the post-pulse incubation time and temperature. Namely, when bacterial cells were incubated 24 h at 37°C, the leakage of bacterial proteins was approximately 2 or 3 – times higher compared to 24 h incubation at room temperature or compared to shorter incubation times (1 to 3 hours).
We can conclude that an effect of treatment parameters on bacteria membrane permeabilization does not correlate in the same way as on bacteria inactivation and protein extraction. Additional experiments are needed however to confirm these observations.

Poster presentation(s) originating from this mission

Event 1st World Congress on Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental Technologies
Location Portorož/Slovenia
Period

06/09 - 10/09/2015

Authors Sasa Haberl Meglic, Eva Levicnik, Elisa Luengo, Javier Raso, Damijan Miklavcic
Title The effect of temperature on protein extraction by electroporation and on bacterial viability

 


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