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

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          Abstract

          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.

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          The online version of this article (10.1007/s00216-020-02451-9) contains supplementary material, which is available to authorised users.

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          Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels.

          Shota Atsumi, Taizo Hanai, James C Liao (2008)
          Global energy and environmental problems have stimulated increased efforts towards synthesizing biofuels from renewable resources. Compared to the traditional biofuel, ethanol, higher alcohols offer advantages as gasoline substitutes because of their higher energy density and lower hygroscopicity. In addition, branched-chain alcohols have higher octane numbers compared with their straight-chain counterparts. However, these alcohols cannot be synthesized economically using native organisms. Here we present a metabolic engineering approach using Escherichia coli to produce higher alcohols including isobutanol, 1-butanol, 2-methyl-1-butanol, 3-methyl-1-butanol and 2-phenylethanol from glucose, a renewable carbon source. This strategy uses the host's highly active amino acid biosynthetic pathway and diverts its 2-keto acid intermediates for alcohol synthesis. In particular, we have achieved high-yield, high-specificity production of isobutanol from glucose. The strategy enables the exploration of biofuels beyond those naturally accumulated to high quantities in microbial fermentation.
          Article 0 comments Cited 434 times – based on 0 reviews     Review now
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            The production of fine chemicals by biotransformations.

            Adrie J J Straathof, Sven Panke, Andreas Schmid (2002)
            Today, biocatalysis is a standard technology for the production of chemicals. An analysis of 134 industrial biotransformations reveals that hydrolases (44%) and redox biocatalysts (30%) are the most prominent categories. Most products are chiral (89%) and are used as fine chemicals. In the chemical industry, successful product developments involve on average a yield of 78%, a volumetric productivity of 15.5 g/(L.h) and a final product concentration of 108 g/L. By contrast, the pharmaceutical industry focuses on time-to-market. The implications of this for future research and development on biocatalysis are discussed.
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              Continuous Flow Synthesis of Chiral Amines in Organic Solvents: Immobilization of E. coli Cells Containing Both ω-Transaminase and PLP

              Leandro H. Andrade, Timothy Jamison, Wolfgang Kroutil (2014)
              E. coli cells containing overexpressed (R)-selective ω-transaminase and the cofactor PLP were immobilized on methacrylate beads suitable for continuous flow applications. The use of an organic solvent suppresses leaching of PLP from the cells; no additional cofactor was required after setting up the packed-bed reactor containing the biocatalyst (ω-TA-PLP). Non-natural ketone substrates were transformed in flow with excellent enantioselectivity (>99% ee). Features of this novel system include high-throughput (30-60 min residence time), clean production (no quench, workup, or purification required), high enzyme stability (the packed-bed reactor can be continuously operated for 1-10 days), and excellent mass recovery.
              Article 0 comments Cited 20 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Martina Viefhues: viefhues@physik.uni-bielefeld.de
                Journal
                Journal ID (nlm-ta): Anal Bioanal Chem
                Journal ID (iso-abbrev): Anal Bioanal Chem
                Title: Analytical and Bioanalytical Chemistry
                Publisher: Springer Berlin Heidelberg (Berlin/Heidelberg )
                ISSN (Print): 1618-2642
                ISSN (Electronic): 1618-2650
                Publication date (Electronic): 11 March 2020
                Publication date PMC-release: 11 March 2020
                Publication date (Print): 2020
                Volume: 412
                Issue: 16
                Pages: 3925-3933
                Affiliations
                [1 ]GRID grid.7491.b, ISNI 0000 0001 0944 9128, Experimental Biophysics and Applied Nanosciences, Department of Physics, , Bielefeld University, ; 33615 Bielefeld, Germany
                [2 ]GRID grid.7491.b, ISNI 0000 0001 0944 9128, Industrial Organic Chemistry and Biotechnology, Department of Chemistry, , Bielefeld University, ; 33615 Bielefeld, Germany
                [3 ]GRID grid.7491.b, ISNI 0000 0001 0944 9128, Present Address: Fermentation Engineering, Department of Technology, , Bielefeld University, ; 33615 Bielefeld, Germany
                [4 ]GRID grid.7491.b, ISNI 0000 0001 0944 9128, Present Address: Cellular and Molecular Biotechnology, Department of Technology, , Bielefeld University, ; 33615 Bielefeld, Germany
                Article
                Publisher ID: 2451
                DOI: 10.1007/s00216-020-02451-9
                PMC ID: 7235074
                PubMed ID: 32157360
                SO-VID: b9315492-7b24-49ef-a9a7-2944ca915df8
                Copyright © © The Author(s) 2020
                License:

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                Date received : 31 October 2019
                Date revision received : 13 January 2020
                Date accepted : 23 January 2020
                Categories
                Subject: Research Paper
                Custom metadata
                issue-copyright-statement © Springer-Verlag GmbH Germany, part of Springer Nature 2020

                ScienceOpen disciplines: Analytical chemistry
                Keywords: microfluidics,whole-cell biocatalyst,dielectrophoresis analysis,uv-vis absorbance analysis,cofactor leaching
                Data availability:
                ScienceOpen disciplines: Analytical chemistry
                Keywords: microfluidics, whole-cell biocatalyst, dielectrophoresis analysis, uv-vis absorbance analysis, cofactor leaching

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