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Elasto-inertial focusing and particle migration in high aspect ratio microchannels for highthroughput separation

microsystems & Nanoengineering 2024年第3期10卷 381-394页

作者： Selim Tanriverdi Javier Cruz Shahriar Habibi Kasra Amini Martim Costa Fredrik Lundell Gustaf Mårtensson Luca Brandt Outi Tammisola Aman Russom Division of Nanobiotechnology Department of Protein ScienceScience for Life LaboratoryKTH Royal Institute of TechnologySolna 17165Sweden Division of Microsystems Technology Department of Materials Science and EngineeringUppsala UniversityUppsala 75237Sweden FLOW and SeRC(Swedish e-Science Research Centre) Department of Engineering MechanicsRoyal Institute of TechnologyStockholm SE 10044Sweden FLOW and Fluid Physics Laboratory Department of Engineering MechanicsRoyal Institute of TechnologyStockholmSweden Wallenberg Wood Science Center Royal Institute of TechnologyStockholm SE 10044Sweden Department of Energy and Process Engineering Norwegian University of Science and Technology(NTNU)TrondheimNorway AIMES Center for the Advancement of Integrated Medical and Engineering Sciences at Karolinska Institutet and KTH Royal Institute of Technology StockholmSweden

The combination of flow elasticity and inertia has emerged as a viable tool for focusing and manipulating particles using *** there is considerable interest in the field of elasto-inertial microfluidics owing to its potential applications,research on particle focusing has been mostly limited to low Reynolds numbers(Re<1),and particle migration toward equilibrium positions has not been extensively *** this work,we thoroughly studied particle focusing on the dynamic range of flow rates and particle migration using straight microchannels with a single inlet high aspect *** initially explored several parameters that had an impact on particle focusing,such as the particle size,channel dimensions,concentration of viscoelastic fluid,and flow *** experimental work covered a wide range of dimensionless numbers(0.05

关键词： particle separation straight

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Single-cell polymer coating improves the desiccation tolerance of Metarhizium brunneum blastospores

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Biotechnology for the Environment 2024年第1期1卷 1-15页

作者： Dietsch, Robin Jakobs-Schönwandt, Desiree Blöbaum, Luisa Bondzio, Laila Grünberger, Alexander Patel, Anant WG Fermentation and Formulation of Biologicals and Chemicals Faculty of Engineering and Mathematics Hochschule Bielefeld - University of Applied Sciences and Arts Bielefeld Germany Bioengineering and Sustainability Westphalian University of Applied Sciences Recklinghausen Germany Multiscale Bioengineering Technical Faculty Bielefeld University Bielefeld Germany Thin Films and Physics of Nanostructures Faculty of Physics Bielefeld University Bielefeld Germany Institute of Process Engineering in Life Sciences: Microsystems in Bioprocess Engineering Karlsruhe Institute of Technology Karlsruhe Germany

Equipping cells with artificial shells or coats has been explored throughout the last decade, with goals such as immunomasking, in vivo tracing, and imparting tolerance to various biotic and abiotic stressors. One stressor, however, drying, has curiously been overlooked. In an industrial setting, the drying of cells is relevant when a satisfactory product shelf life must be achieved at a low cost. The drying of entomopathogenic organisms for biocontrol is a prime example of this. Here, the thin-walled blastospores of the entomopathogenic fungus Metarhizium brunneum are a great model organism for testing whether thin-cell surface polyelectrolyte layers may increase desiccation tolerance. In this study, we coated single M. brunneum blastospores with alternating layers of chitosan and alginate and assessed their effects on blastospore viability after drying. The desiccation tolerance improved with increasing layer numbers from 6.9% to a maximum of 27.5%. In addition, as the polymer chain length decreased, the desiccation tolerance further increased to 33.1%. Furthermore, we provided visual proof of the coating surrounding the blastospores via the use of fluorescent polymers and scanning electron microscopy. Finally, an investigation of differences in water absorption into coated and uncoated cells revealed that water absorbed faster into coated cells when alginate was on the surface of the structure but slower when the outermost layer was composed of chitosan. We conclude that, via polyelectrolyte multilayering on thin-walled blastospores, desiccation tolerance can be significantly increased, but a deeper understanding is necessary to extract the full potential from this technique.

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Protocol for microfluidic single-cell cultivation and live-cell imaging of Chinese hamster ovary suspension cell lines

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STAR Protocols 2024年第2期5卷 103106页

作者： Schmitz, Julian Yermakov, Boris Grünberger, Alexander Multiscale Bioengineering Faculty of Technology Bielefeld University Universitätsstraße 25 Bielefeld 33615 Germany Institute of Process Engineering in Life Sciences Microsystems in Bioprocess Engineering Karlsruhe Institute of Technology (KIT) Fritz-Haber-Weg 2 Karlsruhe 76131 Germany

Microfluidic single-cell cultivation (MSCC) is a powerful tool for investigating the cellular behavior of various cell types at the single-cell level. Here, we present a protocol specifically developed for the reliable and reproducible MSCC of industrially relevant Chinese hamster ovary (CHO) suspension cell lines. We summarize critical experimental steps from the initial seed train up to the final MSCC experiment, with a special focus on pre-culture management and medium preparation, device inoculation, and the establishment of a constant medium perfusion. © 2024 The Author(s)

关键词： Biotechnology and bioengineering Cell culture Single Cell

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Solute excretion improves trehalose uptake and desiccation tolerance of Metarhizium brunneum blastospores

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Biotechnology for the Environment 2025年第1期2卷 1-17页

作者： Dietsch, Robin Jakobs-Schönwandt, Desiree Blöbaum, Luisa Persicke, Marcus Grünberger, Alexander Patel, Anant WG Fermentation and Formulation of Biologicals and Chemicals Faculty of Engineering and Mathematics Bielefeld Germany Bioengineering and Sustainability Westfälische Hochschule Recklinghausen Germany Multiscale Bioengineering Technical Faculty Bielefeld University Bielefeld Germany OMICS Core Facility – Proteomics Metabolomics Unit (under construction) Bielefeld University Bielefeld Germany Microbial Genomics and Biotechnology Center for Biotechnology Bielefeld University Bielefeld Germany Institute of Process Engineering in Life Sciences: Microsystems in Bioprocess Engineering Karlsruhe Institute of Technology Karlsruhe Germany

In this study, a novel approach was developed to increase the intracellular trehalose concentration in M. brunneum blastospores by uptake from the medium, improving their desiccation tolerance. Briefly, placing the blastospores in a hypotonic environment induced solute excretion, allowing significantly increased trehalose uptake during subsequent trehalose treatment. Conductivity changes and GC–MS analysis revealed that glycerol and lactate excretion is triggered in a hypotonic NaCl solution. After hypotonic + trehalose treatment, intracellular glycerol, lactate, and glucose levels increased, while other desiccation-protective solutes remained unaffected. A fluorescence-based single-cell analysis of membrane stress and death during osmotic desiccation and rehydration showed that membrane stress and cell death were strongly amplified upon rehydration. Trehalose treatment reduced this amplification, while hypotonic + trehalose treatment minimised cell death and prevented a stress response during rehydration, though it increased membrane stress during desiccation. Newly formed germ tubes were identified as critical points of membrane failure during desiccation. Ultimately, the drying survival of blastospores was significantly improved by the hypotonic + trehalose treatment compared to untreated or trehalose-only-treated blastospores. In summary, this technique successfully enhanced blastospore desiccation tolerance and could be adapted to load cells with other solutes.

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Production of Polyhydroxybutyrate from Industrial Flue Gas by Microbial Electrosynthesis

SSRN

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SSRN 2024年

作者： Langsdorf, Alexander Schütz, Julian Philipp Ulber, Roland Stöckl, Markus Holtmann, Dirk Institute of Bioprocess Engineering and Pharmaceutical Technology University of Applied Sciences Mittelhessen Wiesenstrasse 14 Giessen35390 Germany Institute of Bioprocess Engineering University of Kaiserslautern-Landau Gottlieb-Daimler-Straße 49 Kaiserslautern67663 Germany Chemical Technology DECHEMA Research Institute Theodor-Heuss-Allee 25 Frankfurt am Main60486 Germany Institute of Process Engineering in Life Sciences Karlsruhe Institute of Technology Fritz-Haber-Weg 4 Karlsruhe76131 Germany

The aim of this work was to produce the biopolymer polyhydroxybutyrate (PHB) from industrial flue gas as CO2 source and electrolysis originating hydrogen via microbial electrosynthesis. Besides the laboratory experiments, the experiments were carried out under industrial conditions directly on-site in a cogeneration plant. We were able to demonstrate that the use of flue gas as a CO2 source has no detectable negative effect on bacterial growth and PHB production in comparison to a pure gas mixture. In an electrochemical H-cell 333 ± 44 mg L-1 PHB were obtained, which corresponds to a PHB content of 43 ± 3% of the cell dry weight. By using flue gas for the production of the biopolymer PHB, not only CO2 emissions are reduced, but also possible environmental pollution from non-biodegradable plastics. To the best of our knowledge, this is the first time that the production of PHB from flue gas has been demonstrated using Cupriavidus necator. A new production system for various industrially relevant chemicals is possible by coupling the conversion of CO2 from flue gases with water electrolysis as the sole carbon and energy source. © 2024, The Authors. All rights reserved.

关键词： Flue gases

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Application of the All-In-One Electrode for in Situ H2O2 Generation in Hydroxylation Catalyzed by Unspecific Peroxygenase from Agrocybe Aegerita

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SSRN 2023年

作者： Sayoga, Giovanni Vallian Bueschler, Victoria Sophie Beisch, Hubert Holtmann, Dirk Zeng, An-Ping Fiedler, Bodo Ohde, Daniel Liese, Andreas Institute of Technical Biocatalysis Hamburg University of Technology Denickestraße 15 Hamburg21073 Germany Institute of Polymers and Composites Hamburg University of Technology Denickestraße 15 Hamburg21073 Germany Institute of Process Engineering in Life Sciences Karlsruhe Institute of Technology Fritz-Haber-Weg 4 Karlsruhe76131 Germany Institute of Bioprocess and Biosystems Engineering Hamburg University of Technology Denickestraße 15 Hamburg21073 Germany Center of Synthetic Biology and Integrated Bioengineering School of Engineering Westlake University Zhejiang Hangzhou310024 China

The versatile unspecific peroxygenase from the fungus Agrocybe aegerita (AaeUPO) is capable to perform hydroxylation of various substrates, even unactivated C-H bonds. The technical application of AaeUPO is still limited due to an instability towards its co-substrate hydrogen peroxide (H2O2). Electrochemical synthesis of H2O2 is an attractive approach that provides controllable in situ generation of H2O2. The aims of this study are to promote catalyst efficiency and develop an individually tailored system for H2O2-dependent reactions. Herein, an All-in-One electrode (AiO) system with a carbon felt cathode is used to generate H2O2 in situ and combined for the first time with an enzymatic hydroxylation. The AiO electrode combines the counter and working electrode in a single rod structure. This shape provides convenient integration into conventional bioreactors, thus, converting them into bioelectrochemical systems (BES). Hydroxylation of 4-ethylbenzoic acid (EBA) catalyzed by the recombinant AaeUPO (rAaeUPO) was chosen as the model reaction. Total turnover number (TTN) up to 450,000 mol mol-1 and turnover frequency (TOF) up to 7.7 s-1 were achieved using the AiO electrode system. The H2O2 productivity was identified as the limiting factor in the hydroxylation of EBA. However, with numbering-up or surface enlargement, this process could have great potential as an optimizable platform for H2O2-dependent enzymatic reactions. © 2023, The Authors. All rights reserved.

关键词： Hydrogen peroxide

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Iron limitation – A perspective on a growth-restricted cultivation strategy for a H2 production system using the diazotrophic cyanobacterium Nostoc PCC 7120 ΔhupW

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Bioresource Technology Reports 2020年 11卷

作者： Howe, Christoph Becker, Daniel Steinweg, Christian Posten, Clemens Stensjö, Karin Department of Chemistry-Ångström Laboratory Uppsala University Uppsala75120 Sweden Institute of Process Engineering in Life Sciences III Bioprocess Engineering Karlsruhe76131 Germany

H2 is a promising carbon-free alternative to fossil fuels, and can be photoautotrophically produced by cyanobacteria. Despite efforts to increase cyanobacterial H2 production by genetic engineering and technical optimization the yield remains economically non-viable. Recent studies have therefore demanded the impairment of growth to reallocate more carbohydrates into biofuel biosynthesis, and nutrient limitation may be a strategy to achieve that. Here we show that limitation of Fe in the filamentous and heterocystous mutant strain Nostoc PCC 7120 ΔhupW resulted in a 5.3-fold lower chlorophyll a concentration as compared to the controls during four-week cultivation. The Fe-limited filaments contained a heterocyst frequency at ~6% and a 4-fold higher specific carbohydrate concentration as compared to the control cultures. We discuss whether Fe-limiting conditions leading to an accumulation of carbon storage compounds could be used as a heterocyst based H2 production system. © 2020 The Author(s)

关键词： Carbohydrates

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Electrodes from carbonized grass clippings for bioelectrochemical systems

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Cleaner Chemical engineering 2024年 9卷

作者： Alexander Langsdorf Michael Halim Marianne Volkmar Markus Stöckl Ralf Harnisch Peter Hahn Roland Ulber Dirk Holtmann Institute of Bioprocess Engineering and Pharmaceutical Technology University of Applied Sciences Mittelhessen Wiesenstrasse 14 35390 Giessen Germany Institute of Bioprocess Engineering University of Kaiserslautern-Landau Gottlieb-Daimler-Straße 49 67663 Kaiserslautern Germany Chemical Technology DECHEMA Research Institute Theodor-Heuss-Allee 25 60486 Frankfurt am Main Germany ifn Forschungs- und Technologiezentrum GmbH Dr.-Bergius-Straße 19 06729 Elsteraue Germany Institute of Process Engineering in Life Sciences Karlsruhe Institute of Technology Fritz-Haber-Weg 4 76131 Karlsruhe Germany

One major obstacle to the commercialization of electrobiotechnological systems is the cost of materials, including expensive electrodes. Smart recycling as well as the use of renewable resources can contribute to producing electrodes more ecologically and economically. Green waste is a biogenic residual material that occurs mainly in urban areas and is currently not recycled to a sufficient extent. Here we show the fabrication of electrodes from carbonized grass clippings and their application in microbial electrosynthesis as well as microbial fuel cells. While the electrodes cannot compete with established metal competitors for water electrolysis in microbial electrosynthesis, they perform comparably to commercial graphite electrodes in microbial fuel cells. With Geobacter sulfurreducens , a current response can be recorded for more than six weeks. To the best of our knowledge, this is the first time that carbonized green waste has been used as an electrode material for bioelectrochemical systems. This demonstrates the potential of carbonized biological materials as a raw material for electrodes and presents a recycling alternative for green waste.

关键词： Green waste Biochar Electrode Microbial electrosynthesis Microbial fuel cell Biorefinery

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Membrane‐harvesting for optimized wastewater treatment using microalgae

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Chemie Ingenieur Technik 2020年第9期92卷

作者： F. Ortiz Tena C. Steinweg C. Posten Karlsruhe Institute for Technology Institute of Process Engineering in Life Sciences – Bioprocess Engineering Fritz-Haber-Weg 2 76131 Karlsruhe Germany

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Environmental considerations and current status of grouping and regulation of engineered nanomaterials

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Environmental Nanotechnology, Monitoring and Management 2022年 18卷

作者： Tschiche, Harald R. Bierkandt, Frank S. Creutzenberg, Otto Fessard, Valerie Franz, Roland Giese, Bernd Greiner, Ralf Haas, Karl‑Heinz Haase, Andrea Hartwig, Andrea Hund‑Rinke, Kerstin Iden, Pauline Kromer, Charlotte Loeschner, Katrin Mutz, Diana Rakow, Anastasia Rasmussen, Kirsten Rauscher, Hubert Richter, Hannes Schoon, Janosch Schmid, Otmar Som, Claudia M.Tovar, Günter E. Westerhoff, Paul Wohlleben, Wendel Luch, Andreas Laux, Peter German Federal Institute for Risk Assessment (BfR) Department of Chemical and Product Safety Berlin Germany Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM) Hannover Germany French Agency for Food Environmental and Occupational Health & Safety (ANSES) Fougères Laboratory Toxicology of contaminants Unit Fougères France Fraunhofer Institute for Process Engineering and Packaging (IVV) Freising Germany University of Natural Resources and Life Sciences (BOKU) Institute of Safety and Risk Sciences (ISR) Vienna Austria Max Rubner-Institut Department of Food Technology and Bioprocess Engineering Karlsruhe Germany Fraunhofer Institute for Silicate Research (ISC) Würzburg Germany Karlsruhe Institute of Technology (KIT) Institute of Applied Biosciences (IAB) Food Chemistry and Toxicology Germany Fraunhofer Institute for Molecular Biology and Applied Ecology (IME) Schmallenberg Germany Nanid Scientific Consulting Germany National Food Institute Technical University of Denmark Lyngby Denmark German Federal Institute for Risk Assessment (BfR) Research Strategy and Coordination Berlin Germany Charité - Universitätsmedizin Berlin Center for Musculoskeletal Surgery Berlin Germany University Medicine Greifswald Center for Orthopaedics Trauma Surgery and Rehabilitation Medicine Greifswald Germany European Commission Joint Research Centre (JRC) Ispra Italy Fraunhofer IKTS - Institute for Ceramic Technologies and Systems Hermsdorf Germany Berlin Institute of Health at Charité - Universitätsmedizin Berlin Julius Wolff Institute Berlin Germany Comprehensive Pneumology Center (CPC-M) Member of the German Center for Lung Research (DZL) Munich Germany Institute of Lung Health and Immunity Helmholtz Zentrum München – German Research Center for Environmental Health Neuherberg Germany Swiss Federal Laboratories for Materials Science and Technology (Empa) Technology and Society Laboratory St. Gallen Switzerland Fraunhofer Institute for Interfacial Engineering and Biot

This article reviews the current status of nanotechnology with emphasis on application and related environmental considerations as well as legislation. Application and analysis of nanomaterials in infrastructure (construction, building coatings, and water treatment) is discussed, and in particular nanomaterial release during the lifecycle of these applications. Moreover, possible grouping approaches with regard to ecotoxicological and toxicological properties, and the fate of nanomaterials in the environment are evaluated. In terms of potential exposure, the opportunities that arise from leveraging advances in several key areas, such as water treatment and construction are addressed. Additionally, this review describes challenges with regard to the European Commission's definition of ‘nanomaterial’. The revised REACH information requirements, intended to enable a comprehensive risk assessment of nanomaterials, are outlined. © 2022 The Author(s)

关键词： Engineered Nanomaterials Environmental Nanotechnology Grouping Regulation

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