Bioelectrochemistry 业 ELSEVIER Bioelectrochemistry 55(2002)107-112 www.elsevier.com/locate/bioelechem Recent biotechnological developments of electropulsation. A prospective review J.Teissie*,N.Eynard,M.C.Vernhes,A.Benichou,V.Ganeva,B.Galutzovb, P.A.Cabanes "IPBS CNRS (UMR 5089).205 Route de Narbonne,31077 Toulouse Cedex.France bUniversity of Sofia,Sofia.Bulgaria SEM EDF Paris,France Received 31 July 2001;received in revised form 12 October 2001;accepted 15 October 2001 Abstract During the last 25 years,basic research has improved our knowledge on the molecular mechanisms triggered at the membrane level by electric pulses.Applied aspects may now be used under safe conditions. Electropulsation is known as a very efficient tool for obtaining gene transfer in many species to produce genetically modified organisms (GMO).This is routinely used for industrial purposes to transfer exogenous activities in bacteria,yeasts and plants.The method is simple and of a low cost. But electropulsation is not limited to this application for biotechnological purposes.It is known that the field-associated membrane alterations can be irreversible.The pulsed species cannot recover after the treatment.Their viability is strongly affected.This appears as a very promising technology for the eradication of pathogenic microorganisms.Recent developments are proposed for sterilization purposes. New flow technologies of field generation allow the treatment of large volumes of solution.When high flow rates are used,microorganisms are submitted both to a hydromechanical and to an electrical stress.The synergy of the two effects may be present when suitable pulsing conditions are chosen.Several examples for the treatment of domestic water and in the food industry are described. Walled microorganisms are affected not only at the membrane level.We observed that alterations are present on the cell wall.A very promising technology is the associated controlled leakage of the cytoplasmic soluble proteins.Large dimeric proteins such as B- galactosidases can be extracted at a high yield.High volumes can be treated by using a flow process.Extraction of proteins is obtained with many systems including mammalian cells.2002 Elsevier Science B.V.All rights reserved. Keywords:Electropulsation;Biotechnology;Microorganisms;Yeast;Amoebae;Legionella 1.Introduction use in other biotechnological fields is somewhat limited with the exception of its key function in animal cloning [2]. When during the Szeged BES meeting in 1987,the Electrotransformation (electrically mediated gene transfer) principal investigator was asked to give a survey of the is routinely used at the bench to obtain genetically modified effects of electric fields and currents on living cells and their organisms (GMO)[3-5]. potential use in biotechnology,two main emerging fields But no significant progress at the industrial scale has were present [1].Taking benefit of the basic research results been made to obtain by such technologies: of the early 1980s,electrofusion and electrotransformation were starting to be used not only in academic laboratories (a)an increase in the growth of the microorganism while cell electroactivation was expected to be a fruitful (b)an increase in the metabolic activity of micro- technology.More than 10 years later,while electrofusion is organisms used for the formation of plant hybrids rather routinely,its while this was expected in 1987. In fact biotechnological applications remain focused on ”Corresponding author..Tel:+33-5-6117-5812;fax:+33-5-6117-5994. small-scale experiments.Getting a limited number of trans- E-mail address:justin@ipbs.fr (J.Teissie). formed microorganisms is enough to prepare the availability 1567-5394/02/S-see front matter 2002 Elsevier Science B.V.All rights reserved. P正：S1567-5394(01)00138-4Recent biotechnological developments of electropulsation. A prospective review J. Teissie´ a,*, N. Eynard a , M.C. Vernhes a , A. Be´nichou a , V. Ganeva b , B. Galutzov b , P.A. Cabanes c a IPBS CNRS (UMR 5089), 205 Route de Narbonne, 31077 Toulouse Cedex, France b University of Sofia, Sofia, Bulgaria c SEM EDF, Paris, France Received 31 July 2001; received in revised form 12 October 2001; accepted 15 October 2001 Abstract During the last 25 years, basic research has improved our knowledge on the molecular mechanisms triggered at the membrane level by electric pulses. Applied aspects may now be used under safe conditions. Electropulsation is known as a very efficient tool for obtaining gene transfer in many species to produce genetically modified organisms (GMO). This is routinely used for industrial purposes to transfer exogenous activities in bacteria, yeasts and plants. The method is simple and of a low cost. But electropulsation is not limited to this application for biotechnological purposes. It is known that the field-associated membrane alterations can be irreversible. The pulsed species cannot recover after the treatment. Their viability is strongly affected. This appears as a very promising technology for the eradication of pathogenic microorganisms. Recent developments are proposed for sterilization purposes. New flow technologies of field generation allow the treatment of large volumes of solution. When high flow rates are used, microorganisms are submitted both to a hydromechanical and to an electrical stress. The synergy of the two effects may be present when suitable pulsing conditions are chosen. Several examples for the treatment of domestic water and in the food industry are described. Walled microorganisms are affected not only at the membrane level. We observed that alterations are present on the cell wall. A very promising technology is the associated controlled leakage of the cytoplasmic soluble proteins. Large dimeric proteins such as b￾galactosidases can be extracted at a high yield. High volumes can be treated by using a flow process. Extraction of proteins is obtained with many systems including mammalian cells. D 2002 Elsevier Science B.V. All rights reserved. Keywords: Electropulsation; Biotechnology; Microorganisms; Yeast; Amoebae; Legionella 1. Introduction When during the Szeged BES meeting in 1987, the principal investigator was asked to give a survey of the effects of electric fields and currents on living cells and their potential use in biotechnology, two main emerging fields were present [1]. Taking benefit of the basic research results of the early 1980s, electrofusion and electrotransformation were starting to be used not only in academic laboratories while cell electroactivation was expected to be a fruitful technology. More than 10 years later, while electrofusion is used for the formation of plant hybrids rather routinely, its use in other biotechnological fields is somewhat limited with the exception of its key function in animal cloning [2]. Electrotransformation (electrically mediated gene transfer) is routinely used at the bench to obtain genetically modified organisms (GMO) [3– 5]. But no significant progress at the industrial scale has been made to obtain by such technologies: (a) an increase in the growth of the microorganism (b) an increase in the metabolic activity of micro￾organisms while this was expected in 1987. In fact biotechnological applications remain focused on small-scale experiments. Getting a limited number of trans￾formed microorganisms is enough to prepare the availability 1567-5394/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved. PII: S1567-5394(01)00138-4 * Corresponding author. Tel.: +33-5-6117-5812; fax: +33-5-6117-5994. E-mail address: justin@ipbs.fr (J. Teissie´). www.elsevier.com/locate/bioelechem Bioelectrochemistry 55 (2002) 107 – 112