A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes

In the course of their development, industrial biocatalysis processes have to be optimized in small-scale, e. g., within microfluidic bioreactors. Recently, we introduced a novel microfluidic reactor device, which can handle defined reaction compartments of up to 250 μL in combination with magnetic...

Full description

Bibliographic Details
Main Authors: Raphael Heinzler, Jonas Hübner, Thomas Fischöder, Lothar Elling, Matthias Franzreb
Format: Article
Language:English
Published: Frontiers Media S.A. 2018-12-01
Series:Frontiers in Bioengineering and Biotechnology
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fbioe.2018.00189/full
_version_ 1819023815114489856
author Raphael Heinzler
Jonas Hübner
Thomas Fischöder
Lothar Elling
Matthias Franzreb
author_facet Raphael Heinzler
Jonas Hübner
Thomas Fischöder
Lothar Elling
Matthias Franzreb
author_sort Raphael Heinzler
collection DOAJ
description In the course of their development, industrial biocatalysis processes have to be optimized in small-scale, e. g., within microfluidic bioreactors. Recently, we introduced a novel microfluidic reactor device, which can handle defined reaction compartments of up to 250 μL in combination with magnetic micro carriers. By transferring the magnetic carriers between subsequent compartments of differing compositions, small scale synthesis, and bioanalytical assays can be conducted. In the current work, this device is modified and extended to broaden its application range to the screening and optimization of bioprocesses applying immobilized enzymes. Besides scaling the maximum compartment volume up to 3 mL, a temperature control module, as well as a focused infrared spot were integrated. By adjusting the pump rate, compartment volumes can be accurately dosed with an error <5% and adjusted to the requested temperature within less than a minute. For demonstration of bioprocess parameter optimization within such compartments, the influence of pH, temperature, substrate concentration, and enzyme carrier loading was automatically screened for the case of transferring UDP-Gal onto N-acetylglucosamine linked to a tert-butyloxycarbonyl protected amino group using immobilized β1,4-galactosyltransferase-1. In addition, multiple recycling of the enzyme carriers and the use of increased compartment volumes also allows the simple production of preparative amounts of reaction products.
first_indexed 2024-12-21T04:44:53Z
format Article
id doaj.art-a981b1a323d14094991dd2352257e5b6
institution Directory Open Access Journal
issn 2296-4185
language English
last_indexed 2024-12-21T04:44:53Z
publishDate 2018-12-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Bioengineering and Biotechnology
spelling doaj.art-a981b1a323d14094991dd2352257e5b62022-12-21T19:15:36ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852018-12-01610.3389/fbioe.2018.00189413008A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized EnzymesRaphael Heinzler0Jonas Hübner1Thomas Fischöder2Lothar Elling3Matthias Franzreb4Institute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, GermanyInstitute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, GermanyLaboratory for Biomaterials, Institute for Biotechnology and Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, GermanyLaboratory for Biomaterials, Institute for Biotechnology and Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, GermanyInstitute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, GermanyIn the course of their development, industrial biocatalysis processes have to be optimized in small-scale, e. g., within microfluidic bioreactors. Recently, we introduced a novel microfluidic reactor device, which can handle defined reaction compartments of up to 250 μL in combination with magnetic micro carriers. By transferring the magnetic carriers between subsequent compartments of differing compositions, small scale synthesis, and bioanalytical assays can be conducted. In the current work, this device is modified and extended to broaden its application range to the screening and optimization of bioprocesses applying immobilized enzymes. Besides scaling the maximum compartment volume up to 3 mL, a temperature control module, as well as a focused infrared spot were integrated. By adjusting the pump rate, compartment volumes can be accurately dosed with an error <5% and adjusted to the requested temperature within less than a minute. For demonstration of bioprocess parameter optimization within such compartments, the influence of pH, temperature, substrate concentration, and enzyme carrier loading was automatically screened for the case of transferring UDP-Gal onto N-acetylglucosamine linked to a tert-butyloxycarbonyl protected amino group using immobilized β1,4-galactosyltransferase-1. In addition, multiple recycling of the enzyme carriers and the use of increased compartment volumes also allows the simple production of preparative amounts of reaction products.https://www.frontiersin.org/article/10.3389/fbioe.2018.00189/fullbioreactormagnetic particlesprocess development and optimizationimmobilized enzymatic reactorautomated microreaction system
spellingShingle Raphael Heinzler
Jonas Hübner
Thomas Fischöder
Lothar Elling
Matthias Franzreb
A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes
Frontiers in Bioengineering and Biotechnology
bioreactor
magnetic particles
process development and optimization
immobilized enzymatic reactor
automated microreaction system
title A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes
title_full A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes
title_fullStr A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes
title_full_unstemmed A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes
title_short A Compartmented Flow Microreactor System for Automated Optimization of Bioprocesses Applying Immobilized Enzymes
title_sort compartmented flow microreactor system for automated optimization of bioprocesses applying immobilized enzymes
topic bioreactor
magnetic particles
process development and optimization
immobilized enzymatic reactor
automated microreaction system
url https://www.frontiersin.org/article/10.3389/fbioe.2018.00189/full
work_keys_str_mv AT raphaelheinzler acompartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT jonashubner acompartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT thomasfischoder acompartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT lotharelling acompartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT matthiasfranzreb acompartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT raphaelheinzler compartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT jonashubner compartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT thomasfischoder compartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT lotharelling compartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes
AT matthiasfranzreb compartmentedflowmicroreactorsystemforautomatedoptimizationofbioprocessesapplyingimmobilizedenzymes