Dielectrophoretic analysis of the impact of isopropyl alcohol on the electric polarisability of Escherichia coli whole-cells

Autor:	Miriam S. Epping, Anke Hummel, Marco T. Radukic, Martina Viefhues, Severin Wedde, Armin Grundmann, Harald Gröger
Rok vydání:	2020
Předmět:	Electrophoresis Microfluidics 02 engineering and technology Whole-cell biocatalyst medicine.disease_cause 01 natural sciences Biochemistry Cofactor Analytical Chemistry 2-Propanol Absorbance chemistry.chemical_compound Escherichia coli medicine Solubility Aqueous solution biology Chemistry 010401 analytical chemistry UV-Vis absorbance analysis Isopropyl alcohol Dielectrophoresis 021001 nanoscience & nanotechnology Combinatorial chemistry Culture Media 0104 chemical sciences Dielectrophoresis analysis Biocatalysis Solvents biology.protein NAD+ kinase Cofactor leaching 0210 nano-technology Research Paper
Zdroj:	Analytical and Bioanalytical Chemistry
ISSN:	1618-2650 1618-2642
Popis:	Whole-cell biocatalysts are versatile tools in (industrial) production processes; though, the effects that impact the efficiency are not fully understood yet. One main factor that affects whole-cell biocatalysts is the surrounding medium, which often consists of organic solvents due to low solubility of substrates in aqueous solutions. It is expected that organic solvents change the biophysical and biochemical properties of the whole-cell biocatalysts, e.g. by permeabilising the cell membrane, and thus analysis of these effects is of high importance. In this work, we present an analysis method to study the impact of organic solvents on whole-cell biocatalysts by means of dielectrophoresis. For instance, we evaluate the changes of the characteristic dielectrophoretic trapping ratio induced by incubation of Escherichia coli, serving as a model system, in an aqueous medium containing isopropyl alcohol. Therefore, we could evaluate the impact on the electric polarisability of the cells. For this purpose, a special microchannel device was designed and Escherichia coli cells were genetically modified to reliably synthesise a green fluorescent protein. We could demonstrate that our method was capable of revealing different responses to small changes in isopropyl alcohol concentration and incubation duration. Complementary spectrophotometric UV-Vis (ultraviolet-visible light) absorbance analysis of released NAD(P)+/NAD(P)H cofactor and proteins confirmed our results. Based on our results, we discuss the biophysical effects taking place during incubation. Graphical abstract Electronic supplementary material The online version of this article (10.1007/s00216-020-02451-9) contains supplementary material, which is available to authorised users.
Databáze:	OpenAIRE
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