Microfluidic Irreversible Electroporation-A Versatile Tool to Extract Intracellular Contents of Bacteria and Yeast.

Autor:	Rockenbach A; Institute of Materials in Electrical Engineering 1, RWTH Aachen, 52074 Aachen, Germany. Alexander.Rockenbach@rwth-aachen.de., Sudarsan S; iAMB Institute of Applied Microbiology, ABBt-Aachen Biology and Biotechnology Department, RWTH Aachen, 52074 Aachen, Germany. sursud@biosustain.dtu.dk.; Current address: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark. sursud@biosustain.dtu.dk., Berens J; Institute of Materials in Electrical Engineering 1, RWTH Aachen, 52074 Aachen, Germany. judith.berens@rwth-aachen.de., Kosubek M; Institute of Materials in Electrical Engineering 1, RWTH Aachen, 52074 Aachen, Germany. Kosubek.m92@googlemail.com., Lazar J; Institute of Materials in Electrical Engineering 1, RWTH Aachen, 52074 Aachen, Germany. jaroslav.lazar@rwth-aachen.de., Demling P; iAMB Institute of Applied Microbiology, ABBt-Aachen Biology and Biotechnology Department, RWTH Aachen, 52074 Aachen, Germany. philipp.demling@rwth-aachen.de., Hanke R; iAMB Institute of Applied Microbiology, ABBt-Aachen Biology and Biotechnology Department, RWTH Aachen, 52074 Aachen, Germany. rene.hanke@rwth-aachen.de., Mennicken P; iAMB Institute of Applied Microbiology, ABBt-Aachen Biology and Biotechnology Department, RWTH Aachen, 52074 Aachen, Germany. philip.mennicken@rwth-aachen.de., Ebert BE; iAMB Institute of Applied Microbiology, ABBt-Aachen Biology and Biotechnology Department, RWTH Aachen, 52074 Aachen, Germany. birgitta.ebert@uq.edu.au.; Current address: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, 4072 Brisbane, QLD, Australia. birgitta.ebert@uq.edu.au., Blank LM; iAMB Institute of Applied Microbiology, ABBt-Aachen Biology and Biotechnology Department, RWTH Aachen, 52074 Aachen, Germany. lars.blank@rwth-aachen.de.; Bioeconomy Science Center BioSC, c/o Forschungszentrum Jülich, 52425 Jülich, Germany. lars.blank@rwth-aachen.de., Schnakenberg U; Institute of Materials in Electrical Engineering 1, RWTH Aachen, 52074 Aachen, Germany. schnakenberg@iwe1.rwth-aachen.de.
Jazyk:	angličtina
Zdroj:	Metabolites [Metabolites] 2019 Sep 30; Vol. 9 (10). Date of Electronic Publication: 2019 Sep 30.
DOI:	10.3390/metabo9100211
Abstrakt:	Exploring the dynamic behavior of cellular metabolism requires a standard laboratory method that guarantees rapid sampling and extraction of the cellular content. We propose a versatile sampling technique applicable to cells with different cell wall and cell membrane properties. The technique is based on irreversible electroporation with simultaneous quenching and extraction by using a microfluidic device. By application of electric pulses in the millisecond range, permanent lethal pores are formed in the cell membrane of Escherichia coli and Saccharomyces cerevisiae , facilitating the release of the cellular contents; here demonstrated by the measurement of glucose-6-phosphate and the activity of the enzyme glucose-6-phosphate dehydrogenase. The successful application of this device was demonstrated by pulsed electric field treatment in a flow-through configuration of the microfluidic chip in combination with sampling, inactivation, and extraction of the intracellular content in a few seconds. Minimum electric field strengths of 10 kV/cm for E. coli and 7.5 kV/cm for yeast S. cerevisiae were required for successful cell lysis. The results are discussed in the context of applications in industrial biotechnology, where metabolomics analyses are important.
Databáze:	MEDLINE
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