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Micromachines
Special Issues
Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019)
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Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019)

A special issue of Micromachines (ISSN 2072-666X).

Deadline for manuscript submissions: closed (1 October 2019) | Viewed by 23633

Special Issue Editors

Prof. Dr. Polona Žnidaršič-Plazl

E-Mail Website
Guest Editor
Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia
Interests: biotransformations; microreactors; bioprocess integration
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Bruno Zelić

E-Mail Website
Guest Editor
Faculty of Chemical Engineering and Technology, University of Zagreb, HR-10000 Zagreb, Croatia
Interests: microreactors and microdevices; biodiesel; biogas; downstream processes; solid state fermentation; fermentation; enzymes; enzyme kinetics; mathematical modeling
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The 5th International Conference Implementation of Microreactor Technology in Biotechnology – IMTB 2019 – will take place in Cavtat, Croatia, from 19th to 22nd May 2019. The IMTB conference aims to provide a platform for people from industry and academia, working in the cross-section of microfluidics, life sciences, analytics, and bioprocess engineering. The interdisciplinary feature of the IMTB conferences enables participants to present and discuss their latest results, developments, and strategies within these complementary fields and to integrate them to gain new accomplishments for industrial implementation.

We welcome manuscripts on any subtopic in this area, including:

  1. Enzymatic microreactors (multiphase microflow systems, enzyme immobilization in microreactors, kinetic studies and modeling of enzymatic reactions in microreactors, multistep enzymatic reactions in microflow systems)
  2. Cells within microdevices (microbioreactors for cell culturing, whole-cell biotransformations within microreactors, microfluidic high-throughput screening, stem cells cultivation in microreactors, cell analysis within microdevices)
  3. Analytical microdevices (analysis of biomolecules in microdevices, monitoring within microflow systems, medical diagnostics in microdevices, microarrays, μTAS)
  4. Bioprocess intensification and integration (bioprocess intensification, downstream processing in microflow systems, continuous bioprocess integration, lab-on-a-chip, microscale-based bioprocess modeling, microreactor technology across the scales).

We hope that this Special Issue will further promote the implementation of microflow systems in biotechnology and especially in industrial applications.

Prof. Dr. Polona Žnidaršič-Plazl
Prof. Dr. Bruno Zelić
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Micromachines is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Enzymatic microreactors
  • cells within microdevices
  • analytical microdevices
  • bioprocess intensification and integration

Published Papers (6 papers)

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Research

12 pages, 1524 KiB  
Article
Copolymeric Hydrogel-Based Immobilization of Yeast Cells for Continuous Biotransformation of Fumaric Acid in a Microreactor
by Tadej Menegatti and Polona Žnidaršič-Plazl
Micromachines 2019, 10(12), 867; https://doi.org/10.3390/mi10120867 - 10 Dec 2019
Cited by 16 | Viewed by 3472
Abstract
Although enzymatic microbioreactors have recently gained lots of attention, reports on the use of whole cells as biocatalysts in microreactors have been rather modest. In this work, an efficient microreactor with permeabilized Saccharomyces cerevisiae cells was developed and used for continuous biotransformation of [...] Read more.
Although enzymatic microbioreactors have recently gained lots of attention, reports on the use of whole cells as biocatalysts in microreactors have been rather modest. In this work, an efficient microreactor with permeabilized Saccharomyces cerevisiae cells was developed and used for continuous biotransformation of fumaric into industrially relevant L-malic acid. The immobilization of yeast cells was achieved by entrapment in a porous structure of various hydrogels. Copolymers based on different ratios of sodium alginate (SA) and polyvinyl alcohol (PVA) were used for hydrogel formation, while calcium chloride and boric or phenylboronic acid were tested as crosslinking agents for SA and PVA, respectively. The influence of hydrogel composition on physico-chemical properties of hydrogels prepared in the form of thin films was evaluated. Immobilization of permeabilized S. cerevisiae cells in the selected copolymeric hydrogel resulted in up to 72% retained fumarase activity. The continuous biotransformation process using two layers of hydrogels integrated into a two-plate microreactor revealed high space time yield of 2.86 g/(L·h) while no activity loss was recorded during 7 days of continuous operation. Full article
(This article belongs to the Special Issue Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019))
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12 pages, 3303 KiB  
Article
A Phenolic Acid Decarboxylase-Based All-Enzyme Hydrogel for Flow Reactor Technology
by Esther Mittmann, Sabrina Gallus, Patrick Bitterwolf, Claude Oelschlaeger, Norbert Willenbacher, Christof M. Niemeyer and Kersten S. Rabe
Micromachines 2019, 10(12), 795; https://doi.org/10.3390/mi10120795 - 20 Nov 2019
Cited by 26 | Viewed by 4544
Abstract
Carrier-free enzyme immobilization techniques are an important development in the field of efficient and streamlined continuous synthetic processes using microreactors. Here, the use of monolithic, self-assembling all-enzyme hydrogels is expanded to phenolic acid decarboxylases. This provides access to the continuous flow production of [...] Read more.
Carrier-free enzyme immobilization techniques are an important development in the field of efficient and streamlined continuous synthetic processes using microreactors. Here, the use of monolithic, self-assembling all-enzyme hydrogels is expanded to phenolic acid decarboxylases. This provides access to the continuous flow production of p-hydroxystyrene from p-coumaric acid for more than 10 h with conversions ≥98% and space time yields of 57.7 g·(d·L)−1. Furthermore, modulation of the degree of crosslinking in the hydrogels resulted in a defined variation of the rheological behavior in terms of elasticity and mesh size of the corresponding materials. This work is addressing the demand of sustainable strategies for defunctionalization of renewable feedstocks. Full article
(This article belongs to the Special Issue Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019))
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9 pages, 5211 KiB  
Article
Imine Reductase Based All-Enzyme Hydrogel with Intrinsic Cofactor Regeneration for Flow Biocatalysis
by Patrick Bitterwolf, Felix Ott, Kersten S. Rabe and Christof M. Niemeyer
Micromachines 2019, 10(11), 783; https://doi.org/10.3390/mi10110783 - 15 Nov 2019
Cited by 19 | Viewed by 4153
Abstract
All-enzyme hydrogels are biocatalytic materials, with which various enzymes can be immobilized in microreactors in a simple, mild, and efficient manner to be used for continuous flow processes. Here we present the construction and application of a cofactor regenerating hydrogel based on the [...] Read more.
All-enzyme hydrogels are biocatalytic materials, with which various enzymes can be immobilized in microreactors in a simple, mild, and efficient manner to be used for continuous flow processes. Here we present the construction and application of a cofactor regenerating hydrogel based on the imine reductase GF3546 from Streptomyces sp. combined with the cofactor regenerating glucose-1-dehydrogenase from Bacillus subtilis. The resulting hydrogel materials were characterized in terms of binding kinetics and viscoelastic properties. The materials were formed by rapid covalent crosslinking in less than 5 min, and they showed a typical mesh size of 67 ± 2 nm. The gels were applied for continuous flow biocatalysis. In a microfluidic reactor setup, the hydrogels showed excellent conversions of imines to amines for up to 40 h in continuous flow mode. Variation of flow rates led to a process where the gels showed a maximum space-time-yield of 150 g·(L·day)−1 at 100 μL/min. Full article
(This article belongs to the Special Issue Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019))
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18 pages, 2394 KiB  
Article
Kinetic Parameter Estimation and Mathematical Modelling of Lipase Catalysed Biodiesel Synthesis in a Microreactor
by Martin Gojun, Lucija Pustahija, Ana Jurinjak Tušek, Anita Šalić, Davor Valinger and Bruno Zelić
Micromachines 2019, 10(11), 759; https://doi.org/10.3390/mi10110759 - 08 Nov 2019
Cited by 17 | Viewed by 3136
Abstract
Development of green, clean, and sustainable processes presents new challenges in today’s science. Production of fuel is no exception. Considering the utilisation of various renewable sources, the synthesis of biodiesel, characterised as more environmentally-friendly then fossil fuel, has drawn significant attention. Even though [...] Read more.
Development of green, clean, and sustainable processes presents new challenges in today’s science. Production of fuel is no exception. Considering the utilisation of various renewable sources, the synthesis of biodiesel, characterised as more environmentally-friendly then fossil fuel, has drawn significant attention. Even though the process based on chemical transesterification in a batch reactor still presents the most used method for its production, enzyme catalysed synthesis of biodiesel in a microreactor could be a new approach for going green. In this research, edible sunflower oil and methanol were used as substrates and lipase from Thermomyces lanuginosus (Lipolase L100) was used as catalyst for biodiesel synthesis. Experiments were performed in a polytetrafluoroethylene (PTFE) microreactor with three inlets and in glass microreactors with two and three inlets. For a residence time of 32 min, the fatty acids methyl esters (FAME) yield was 30% higher than the yield obtained for the glass microreactor with three inlets. In comparison, when the reaction was performed in a batch reactor (V = 500 mL), the same FAME yield was achieved after 1.5 h. In order to enhance the productivity of the process, we used proposed reaction kinetics, estimated kinetic parameters, and a mathematical model we developed. After validation using independent experimental data, a proposed model was used for process optimization in order to obtain the highest FAME yield for the shortest residence time. Full article
(This article belongs to the Special Issue Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019))
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11 pages, 2705 KiB  
Article
A Passive Microfluidic Device for Chemotaxis Studies
by Maria Laura Coluccio, Maria Antonia D’Attimo, Costanza Maria Cristiani, Patrizio Candeloro, Elvira Parrotta, Elisabetta Dattola, Francesco Guzzi, Giovanni Cuda, Ernesto Lamanna, Ennio Carbone, Ulrich Krühne, Enzo Di Fabrizio and Gerardo Perozziello
Micromachines 2019, 10(8), 551; https://doi.org/10.3390/mi10080551 - 20 Aug 2019
Cited by 19 | Viewed by 4184
Abstract
This work presents a disposable passive microfluidic system, allowing chemotaxis studies, through the generation of a concentration gradient. The device can handle liquid flows without an external supply of pressure or electric gradients, but simply using gravity force. It is able to ensure [...] Read more.
This work presents a disposable passive microfluidic system, allowing chemotaxis studies, through the generation of a concentration gradient. The device can handle liquid flows without an external supply of pressure or electric gradients, but simply using gravity force. It is able to ensure flow rates of 10 µL/h decreasing linearly with 2.5% in 24 h. The device is made of poly(methylmethacrylate) (PMMA), a biocompatible material, and it is fabricated by micro-milling and solvent assisted bonding. It is assembled into a mini incubator, designed properly for cell biology studies in passive microfluidic devices, which provides control of temperature and humidity levels, a contamination-free environment for cells with air and 5% of CO2. Furthermore, the mini incubator can be mounted on standard inverted optical microscopes. By using our microfluidic device integrated into the mini incubator, we are able to evaluate and follow in real-time the migration of any cell line to a chemotactic agent. The device is validated by showing cell migration at a rate of 0.36 µm/min, comparable with the rates present in scientific literature. Full article
(This article belongs to the Special Issue Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019))
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16 pages, 4075 KiB  
Article
Resonant Mixing in Glass Bowl Microbioreactor Investigated by Microparticle Image Velocimetry
by Sven Meinen, Lasse Jannis Frey, Rainer Krull and Andreas Dietzel
Micromachines 2019, 10(5), 284; https://doi.org/10.3390/mi10050284 - 27 Apr 2019
Cited by 7 | Viewed by 3591
Abstract
Microbioreactors are gaining increased interest in biopharmaceutical research. Due to their decreasing size, the parallelization of multiple reactors allows for simultaneous experiments. This enables the generation of high amounts of valuable data with minimal consumption of precious pharmaceutical substances. However, in bioreactors of [...] Read more.
Microbioreactors are gaining increased interest in biopharmaceutical research. Due to their decreasing size, the parallelization of multiple reactors allows for simultaneous experiments. This enables the generation of high amounts of valuable data with minimal consumption of precious pharmaceutical substances. However, in bioreactors of all scales, fast mixing represents a crucial condition. Efficient transportation of nutrients to the cells ensures good growing conditions, homogeneous environmental conditions for all cultivated cells, and therefore reproducible and valid data. For these reasons, a new type of batch microbioreactor was developed in which any moving mixer component is rendered obsolete through the utilization of capillary surface waves for homogenization. The bioreactor was fabricated in photosensitive glass and its fluid volume of up to 8 µL was provided within a bowl-shaped volume. External mechanical actuators excited capillary surface waves and stereo microparticle image velocimetry (µPIV) was used to analyze resulting convection at different excitation conditions in varied reactor geometries. Typical vortex patterns were observed at certain resonance frequencies where best mixing conditions occurred. Based on the results, a simplified 1D model which predicts resonance frequencies was evaluated. Cultivation of Escherichia coli BL21 under various mixing conditions showed that mixing in resonance increased the biomass growth rate, led to high biomass concentrations, and provided favorable growth conditions. Since glass slides containing multiple bowl reactors can be excited as a whole, massive parallelization is foreseen. Full article
(This article belongs to the Special Issue Implementation of Microreactor Technology in Biotechnology 2019 (IMTB 2019))
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