Sascha Beutel

Leibniz Universität Hannover, Hanover, Lower Saxony, Germany

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Publications (77)128.12 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: The fabrication of individual labware is a sophisticated task that requires dedicated machines and skills. 3D-printing has the great potential to drastically simplify this procedure. In the near future scientists will be able to design labware digitally and 3D-print them afterwards directly in the laboratory. With the available rapid prototyping (RP) printer-systems it is possible to achieve this. The materials accessible are able to meet the needs of biotechnological laboratories which include biocompatibility and withstanding sterilization conditions. This will lead to a completely new approach of adapting the labware to the experiment or even tailor-made it to the organism it is being used for, not adapting the experiment to a certain standard labware. Thus it will encourage the creativity of scientists and will enrich the laboratory work of the future. We present different examples illustrating the potential and the possibilities of using 3D-printing for individualizing labware. This includes a well plate with different baffle geometries, a shake flask cap with build in luer connections and a filter holder for an in-house developed membrane reactor-system.This article is protected by copyright. All rights reserved
    Engineering in Life Sciences 07/2014; · 1.63 Impact Factor
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    ABSTRACT: 3D-printing techniques are continuously evolving, thus their application fields are likewise growing very fast. The applications shown here highlight the use of rapid prototyping (RP) in a dedicated biotechnology laboratory environment. The combination of improving prototypes using fused deposition modeling (FDM) printers and producing useable parts with selective laser sintering (SLS) printers enable a cost- and time-efficient use of these techniques. Biocompatible materials for 3D-printing are already available and the printed parts can directly be used in the laboratory. To demonstrate this, we tested 3D-printing materials for their in vitro biocompatibility. To exemplify the versatility of the 3D-printing process applied to a biotechnology laboratory, a normal well plate design was modified in silico to include different baffle geometries. This plate was subsequently 3D-printed and used for cultivation. In the near future this design and print possibility will revolutionize the industry. Advanced printers will become available for laboratories and could be used for creating individual labware or standard disposables on demand. These applications have the potential to change the way research is done and even how stock-keeping management is today, leading to more flexibility and promoting creativity of the scientists.This article is protected by copyright. All rights reserved
    Engineering in Life Sciences 07/2014; · 1.63 Impact Factor
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    ABSTRACT: A novel online sensor system for non-invasive and continuous monitoring of cell growth in shake flasks is described. The measurement principle is based on turbidity measurement by detecting 180°-scattered light and correlation to optical density by non-linear calibration models. The sensor system was integrated into a commercial shaking tablar to read out turbidity from below the shake flasks bottom. The system was evaluated with two model microorganisms – Escherichia coli K12 as prokaryotic and Saccharomyces cerevisiae as eukaryotic model. The sensor allowed an accurate monitoring of turbidity and correlation with OD600 ≤ 30. The determination of online OD showed relative errors of about 7.5% for E. coli K12 and 12% for S. cerevisiae. This matches the errors of the laborious offline OD and thus facilitates to overcome the drawbacks of the classical method as risk of contamination and decreasing volumes through sampling. One major challenge was to ensure a defined, non-varying measurement zone as the rotating suspension in the shake flask forms a liquid sickle which circulates round the flasks inner bottom wall. The resulting alteration of liquid height above the sensor could be compensated by integration of an acceleration sensor into the tablar to synchronize the sensor triggering.This article is protected by copyright. All rights reserved
    Engineering in Life Sciences 06/2014; · 1.63 Impact Factor
  • Anna Glyk, Thomas Scheper, Sascha Beutel
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    ABSTRACT: Different poly(ethylene glycol) (PEG) + potassium phosphate or sodium citrate aqueous two-phase systems (ATPS) were investigated at 23 °C, containing different PEG types (molecular weights 2000 g·mol–1 to 8000 g·mol–1) and pH values (5 to 9). Furthermore, the effect of the added salt NaCl (0 wt % to 8 wt %) on the PEG + potassium phosphate/sodium citrate ATPS was studied at 23 °C. The experimental binodal data were successfully correlated with the empirical nonlinear equation proposed by Hu. The effects of increasing molecular weight of PEG, pH, NaCl, and salt type on the obtained binodal curves were determined, resulting in a binodal curve shift toward the origin. Thus, an expansion of the two-phase region occurred by increasing molecular weight of the PEG, pH, and NaCl and due to the Gibbs free energy of hydration of ions of phosphate. Furthermore, the phase equilibrium compositions, tie-line lengths, slopes of tie-lines, critical points, and effective excluded volumes were obtained for all studied systems. Finally, the experimental tie-line compositions were successfully correlated by using the Othmer–Tobias and Bancroft equations, and linear dependency was confirmed.
    Journal of Chemical & Engineering Data 02/2014; 59(3):850–859. · 2.00 Impact Factor
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    ABSTRACT: Flavonoids are a large group of plant secondary metabolites with a variety of biological properties and are therefore of interest to many scientists, as they can lead to industrially interesting intermediates. The anaerobic gut bacterium Eubacterium ramulus can catabolize flavonoids, but until now, the pathway has not been experimentally confirmed. In the present work, a chalcone isomerase (CHI) and an enoate reductase (ERED) could be identified through whole genome sequencing and gene motif search. These two enzymes were successfully cloned and expressed in Escherichia coli in their active form, even under aerobic conditions. The catabolic pathway of E. ramulus was confirmed by biotransformations of flavanones into dihydrochalcones. The engineered E. coli strain that expresses both enzymes was used for the conversion of several flavanones, underlining the applicability of this biocatalytic cascade reaction.
    Angewandte Chemie International Edition 01/2014; 53(5):1439-42. · 11.34 Impact Factor
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    ABSTRACT: Probing a cDNA library extracted from Pogostemon cablin (Indian Patchouli) with gene specific primers, a variant of patchoulol synthase PTS (GenBank acc. no.: AY508730) was amplified, cloned, and sequenced. The amino acid sequence deduced from the cloned cDNA exhibited a sequence variation of 3.4 % compared to the annotated sequence. The enzyme variant tended to form inclusion bodies when expressed in E. coli. The coding sequence was fused to the T7-tag, His-tag and to thioredoxin. Constructs were expressed in three different E. coli expression strains, with several strain/construct combinations yielding soluble enzyme. By fusion to thioredoxin and careful codon optimization of the eukaryotic sequence, soluble expression could be improved on average by 42% in comparison to an unoptimized, His-tagged construct. The thioredoxin-fused protein was successfully purified using a one-step Co2+-IMAC purification procedure. Bioactivity assays using prepared farnesyl diphosphate (FDP) in milliliter-scale batch reactions, showed activity of the fused enzyme even with thioredoxin attached. The product spectrum of the enzyme was compared to patchouli oil standards by GC-MS and the main products identified. Interestingly, the variant showed a shift in product spectrum with germacrene A being the most abundant product instead of patchouli alcohol. In silico structural modelling shows a possible chemical and structural change in the active site of the enzyme, which might be responsible for the shift in product composition.
    Protein Expression and Purification 01/2014; · 1.43 Impact Factor
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    ABSTRACT: In the context of this work we evaluated a multisensory, noninvasive prototype platform for shake flask cultivations by monitoring three basic parameters (pH, pO2 and biomass). The focus lies on the evaluation of the biomass sensor based on backward light scattering. The application spectrum was expanded to four new organisms in addition to E. coli K12 and S. cerevisiae [1]. It could be shown that the sensor is appropriate for a wide range of standard microorganisms, e.g., L. zeae, K. pastoris, A. niger and CHO-K1. The biomass sensor signal could successfully be correlated and calibrated with well-known measurement methods like OD600, cell dry weight (CDW) and cell concentration. Logarithmic and Bleasdale-Nelder derived functions were adequate for data fitting. Measurements at low cell concentrations proved to be critical in terms of a high signal to noise ratio, but the integration of a custom made light shade in the shake flask improved these measurements significantly. This sensor based measurement method has a high potential to initiate a new generation of online bioprocess monitoring. Metabolic studies will particularly benefit from the multisensory data acquisition. The sensor is already used in labscale experiments for shake flask cultivations.
    Sensors (Basel, Switzerland). 01/2014; 14(9):17390-17405.
  • Anna Glyk, Thomas Scheper, Sascha Beutel
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    ABSTRACT: In the present work, the quantification of different poly(ethylene glycol) (PEG) + potassium phosphate / sodium citrate aqueous two-phase system (ATPS) phase-forming components was investigated by using conductivity and refractive index measurements. For this purpose, refractive index and conductivity calibration curves were obtained for ATPS at different pH values in the presence of different bovine serum albumin (BSA) concentrations. While BSA had no effect on the conductivity, it had a considerable effect on the refractive index. Finally, a convenient dilution of the samples prior to the ATPS constituent determination is needed to ensure no significant influence from BSA.
    Analytical Biochemistry 01/2014; · 2.58 Impact Factor
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    ABSTRACT: Flavonoide stellen eine große Gruppe pflanzlicher Sekundärmetabolite dar und haben eine Vielzahl unterschiedlicher biologischer Eigenschaften. Aufgrund dessen stehen sie im Interesse vieler Wissenschaftler, da sie zu vielen industriell interessanten Verbindungen führen können. Das anaerobe Darmbakterium Eubacterium ramulus kann über einen bislang unbekannten Abbauweg diese Flavonoide verstoffwechseln. Über eine Vollgenom‐Sequenzierung wurden in dieser Arbeit die entscheidenden Enzyme, eine Chalconisomerase (CHI) und eine Enoatreduktase (ERED), identifiziert, kloniert und funktionell sogar unter aeroben Bedingungen exprimiert. Mittels Biotransformation des Flavanons zum Dihydrochalcon konnte der Flavonoid‐Abbauweg bewiesen werden. Der beide Enzyme exprimierende E.‐coli‐Stamm kann für die Umsetzung verschiedener Flavanone eingesetzt werden. Dies unterstreicht die Anwendbarkeit des in dieser Arbeit entwickelten biokatalytischen Systems. GermanDer biokatalytisch‐metabolische Weg für die Umsetzung von Flavonoiden aus Eubacterium ramulus wurde identifiziert. Eine Chalconisomerase und eine Enoatreduktase wurden erfolgreich kloniert und in E. coli exprimiert und unter aeroben Bedingungen eingesetzt, obwohl E. ramulus ein strikt anaerobes Bakterium ist. Der beide Enzyme exprimierende E.‐coli‐Stamm kann für die Umsetzung verschiedener Flavanone eingesetzt werden.
    Angewandte Chemie 01/2014; 126(5).
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    ABSTRACT: Generating biomass or biochemically active substances with constant and high quality and quantity requires continuous control of the biochemical environment within the used bioreactor. Thus, sensors or sensor systems that monitor the conditions during the cultivation are required. Since in most bioprocesses it is indispensable to prevent any contamination of the culture medium a continuously working sensor in these applications must either stand the process of sterilization or be non-invasive. In this work we present first results from a non-invasive analysis of the waste gas of a 2 L bioreactor during the cultivation of a model organism (E. coli strain K12) using a mass spectrometer with atmospheric pressure chemical ionization and a gas chromatographic pre-separation. Based on the used ionization method a detection of volatile organic compounds emitted during the lag-phase or the growth of the bacteria with extremely low detection limits is possible. We repeatedly observed various compounds with an abundance correlated to the growth of the culture which was monitored by further invasive and non-invasive sensors.
    11. Dresdner Sensor-Symposium 2013; 12/2013
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    ABSTRACT: Potato fruit juice (PFJ) is a by-product from industrial starch production. It still contains several valuable components such as amino acids, minerals and proteins. An economic technology for the isolation and purification of different native potato proteins is the ion exchange chromatography, which can be performed either by classical bed chromatography or by membrane adsorption chromatography (MA-IEX). An already published MA-IEX process for the downstreaming of PFJ is based on the following steps: prefiltration/microfiltration, fractionation with MA-IEX, ultra-/diafiltration and finally drying. In order to further minimize process complexity and costs, new MA-IEX-modules were designed and tested in this research project to facilitate the processing of crude, particle-containing solutions using a tangential flow through the membranes. Modules with fleece polymer spacers and extruded polymer spacers, as well as different spacer channel sizes were tested for their binding capacities and their long-term stability. An optimized setup was found for the technical scale. Modules with extruded polymer spacers channel size 250μm show the highest binding capacities (anion exchanger approx. 0.34mg/cm(2), cation exchanger approx. 0.16mg/cm(2)), while the modules with extruded polymer spacers channel size 480μm show the best long-term stability with 23 passes without intermediary cleaning.
    Journal of Biotechnology 09/2013; · 3.18 Impact Factor
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    ABSTRACT: A new inline method for the monitoring of enzymatic hydrolysis of cellulose is described. Using a new in situ microscope prototype the non-invasive determination of particle size distributions was possible. For the automated analysis of the acquired images a new processing algorithm called CelluloseAnalyzer was developed. It enabled tracking of the number of particles and moreover allowed monitoring of the proportions of particle size fractions during the course of enzymatic hydrolysis reactions. Using this technique significant differences between hydrolysis with endoglucanases and cellulase mixtures were observed. Furthermore the in situ microscopy results were compared with results from offline measurements with laser diffraction spectroscopy and gel permeation chromatography.
    Analytical Chemistry 07/2013; · 5.82 Impact Factor
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    ABSTRACT: Technical scale (≥5 l) cultivations of shear stress sensitive microorganisms are often difficult to perform, as common bioreactors are usually designed to maximize the oxygen input into the culture medium. This is achieved by mechanical stirrers, causing high shear stress. Examples for shear stress sensitive microorganisms, for which no specific cultivation systems exist, are many anaerobic bacteria and fungi, such as basidiomycetes. In this work a disposable bag bioreactor developed for cultivation of mammalian cells was investigated to evaluate its potential to cultivate shear stress sensitive anaerobic Eubacterium ramulus and shear stress sensitive basidiomycetes Flammulina velutipes and Pleurotus sapidus. All cultivations were compared with conventional stainless steel stirred tank reactors (STR) cultivations. Good growth of all investigated microorganisms cultivated in the bag reactor was found. E. ramulus showed growth rates of μ=0.56 h(-1) (bag) and μ=0.53 h(-1) (STR). Differences concerning morphology, enzymatic activities and growth in fungal cultivations were observed. In the bag reactor growth in form of small, independent pellets was observed while STR cultivations showed intense aggregation. F. velutipes reached higher biomass concentrations (21.2 g l(-1) DCW vs. 16.8 g l(-1) DCW) and up to 2-fold higher peptidolytic activities in comparison to cell cultivation in stirred tank reactors.
    Journal of Biotechnology 07/2013; · 3.18 Impact Factor
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    ABSTRACT: Microalgal cultivations present challenges for monitoring and process control posed by their large scale and the likelihood that they will be composed of multiple species. Cell concentration is a fundamental parameter in any cultivation but is typically measured using off-line methods that may be time-consuming, laborious, or subject to interferences. Here, an in-situ microscope has been adapted for monitoring microalgal cultivations by adding a flow-through cell and adjusting image-processing algorithms. After installation in the bypass of a photobioreactor, the microscope enabled the continuous, automated acquisition of cell count, cell size, and cell morphology data on-line during cultivation processes over a period of 20 days, without sampling. The flow-through microscope was tested in cultivations of Chlamydomonas reinhardtii and Chlorella vulgaris. Cell concentration measurements were in agreement with off-line optical density measurements for both species. In addition, cell size and morphology distributions were obtained that revealed population shifts during the cultivation of C. vulgaris. This monitoring system thus provides a means to obtain detailed, non-invasive insights of microalgal cultivation processes.
    Algal Research. 07/2013; 2(3):253–257.
  • Steffen Henkel, Sascha Beutel
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    ABSTRACT: Die gängigsten Messmethoden zur Bestimmung des pH-Werts sind elektrochemischer oder optischer Natur. Als Messsysteme sind dabei besonders die Glaselektrode oder aber der pH-Teststreifen jedem Anwender geläufig. Daneben existieren aber auch weitere Messprinzipien für speziellere Anwendungen z. B. zur pH-Bestimmung in Geweben, Pasten oder unter extremen Prozessbedingungen. Zudem gibt es Methoden wie die pH-Messung mithilfe der NMR, die allein aufgrund des apparativen Aufwands über ein Nischendasein nicht herauskommen werden. Die verschiedenen Methoden zur pH-Bestimmung werden vorgestellt und ihre Vor- und Nachteile diskutiert. The most used measuring principles for the determination of the pH are of electrochemical and optical nature. As measuring system glass electrodes and indicator papers are known by every user. Beside this there are further measuring principles for special applications for example the pH determination in tissues, pastes or under extreme process conditions. Furthermore there are methods like NMR for pH-measurement, which will always relegate to a niche existence because of their complex and expensive apparatus. This article gives an overview of the different available methods and discusses potential applications.
    Chemie Ingenieur Technik 06/2013; 85(6). · 0.70 Impact Factor
  • Chemie Ingenieur Technik 04/2013; 85(4, SI):476-483. · 0.70 Impact Factor
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    ABSTRACT: Hohlfasermembrankontaktoren wurden bereits häufig für die physikalische und chemisch beschleunigte Absorption von CO2 getestet. Dabei bieten Kontaktoren aufgrund des hohen Oberfläche/Volumen-Verhältnisses einige Prozessvorteile. Dennoch ist der Massentransport in kommerziell erhältlichen Hohlfasermodulen limitiert. Durch die geringen Innendurchmesser der Fasern und die niedrigen Durchflussgeschwindigkeiten ergibt sich ein laminares Strömungsprofil. Um insbesondere den grenzschichtnahen Stofftransport zu erhöhen, wurden in dieser Arbeit durch neuartige Hohlfasergeometrien gezielt Sekundärströmungen (Dean-Wirbel) erzeugt. Zudem wurde der Einfluss des Helixdurchmessers untersucht. Mithilfe der Filmtheorie und der dimensionslosen Dean-Zahl gelang die Vorhersage der Absorptionsraten sowohl für gerade als auch helikal geformte Hohlfasern. Hollow fiber membrane contactors have been already tested for the physical and chemical absorption of CO2. Due to their large surface to volume ratio contactors are a promising alternative to conventional equipment. Nevertheless, the overall mass transport in commercially available modules is limited. Due to the small inner diameters of the fibers and the respective low cross flow velocities only low Reynolds numbers can be realized, which leads to a laminar flow regime. In this study Dean vortices have been generated to enhance the mass transport in the boundary layer. Also the influence of the helical curvature diameters on the absorption performance has been studied. By means of the film theory and the dimensionless Dean number absorption rates for both straight and helically shaped hollow fibers have been successfully predicted.
    Chemie Ingenieur Technik 04/2013; 85(4). · 0.70 Impact Factor
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    ABSTRACT: This article is an overview of bioreactors using tubular membranes such as hollow fibers or ceramic capillaries for cultivation processes. This diverse group of bioreactor is described here in regard to the membrane materials used, operational modes, and configurations. The typical advantages of this kind of system such as environments with low shear stress together with high cell densities and also disadvantages like poor oxygen supply are summed up. As the usage of tubular membrane bioreactors is not restricted to a certain organism, a brief overview of various applications covering nearly all types of cells from prokaryotic to eukaryotic cells is also given here.
    Applied Microbiology and Biotechnology 12/2012; · 3.81 Impact Factor
  • Source
    A. Glyk, S. Beutel, T. Scheper
    Chemie Ingenieur Technik 08/2012; 84(8). · 0.70 Impact Factor
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    ABSTRACT: This work aimed to study the enhancement of oxygen transfer rates in hollow fiber membrane contactors by applying Dean vortices. Systematic investigations of how the geometric parameters, like curve diameter, helical pitch or geometric shape, and flow rates affect the mass transport were done to assess the potential for membrane contactors. The membrane was a hydrophobic microfiltration hollow fiber (polypropylene). Helical, meander and twisted formed hollow fibers with fiber lengths from 5 to 30 cm and curvature diameters between 5 and 19 mm were constructed. The oxygen transfer rates per membrane area were investigated. The curved hollow fibers show a linear dependence between Dean number and enhancement factor and an increase of transfer rates up to an enhancement factor of 2.4. For the meander shaped fibers a critical Dean number/range between 10–20 was found, where the mass transport enhancement is adjusted rapidly to those of helical hollow fibers. Also the mass transfer in packed membrane modules was investigated, taking into account the theoretical packing densities of the different fiber geometries. It was found that a volumetric enhancement of gas transfer rates for helical and meander formed fibers can only be improved applying very small curvature diameters less than 4 mm.
    Journal of Membrane Science 01/2012; 423:342-347. · 4.09 Impact Factor

Publication Stats

174 Citations
128.12 Total Impact Points

Institutions

  • 2002–2014
    • Leibniz Universität Hannover
      • Institute of Technical Chemistry
      Hanover, Lower Saxony, Germany
  • 2011
    • Helmholtz Centre for Infection Research
      Brunswyck, Lower Saxony, Germany
    • Ege University
      • Department of Biochemistry
      İzmir, Izmir, Turkey