Extracting proteins by means of electroporation from different microorganisms is gaining on its importance, as electroporation is a quick, chemical free and effective method. In process of obtaining proteins form bacterial cells, total cell disintegration needs to be avoided. Namely, complete cell destruction necessitates additional purifying of end product, therefore cost of the process could be high. Therefore our goal was to study different electric pulse parameters in order to extract as much proteins as possible from E. coli bacteria, while preserving bacterial viability. Our results show, that by increasing electric field strength the concentration of extracted proteins increases and viability reduces. The strong correlation was also found with increasing pulse duration. Pulse number and/or pulse repetition frequency at our conditions has no or little effect on concentration of extracted proteins and/or bacterial viability. We can conclude that the most promising pulse protocol for protein extraction by means of electroporation based on our experience would be longer pulses with lower pulse amplitude assuring high protein yield and low effect on bacterial viability.
COBISS.SI-ID: 11153492
Electroporation is already an established technique in several areas of medicine, but many of its biotechnological applications have only started to emerge; we review here some of the most promising. We outline electroporation as a phenomenon and then proceed to applications, first outlining the best established – the use of reversible electroporation for heritable genetic modification of microorganisms (electrotransformation), and then explore recent advances in applying electroporation for inactivation of microorganisms, extraction of biomolecules, and fast drying of biomass. Although these applications often aim to upscale to the industrial and/or clinical level, we also outline some important chip-scale applications of electroporation. We conclude our review with a discussion of the main challenges and future perspectives.
COBISS.SI-ID: 11070548