μMORE: A microfluidic magnetic oscillation reactor for accelerated parameter optimization in biocatalysis.
Enzymatic parameter determination is an essential step in biocatalytic process development. Therefore higher throughput in miniaturized devices is urgently needed. An ideal microfluidic device should combine easy immobilization and retention of a minimal amount of biocatalyst with a well-mixed reaction volume. Together, all criteria are hardly met by current tools. Here we describe a microfluidic reactor (μMORE) which employs magnetic particles for both enzyme immobilization and efficient mixing using two permanent magnets placed in rotating cylinders next to the a glass chip reactor. The chip geometry and agitation speed was optimized by investigation of the mixing and retention characteristics using simulation and dye distribution analysis. Subsequently, the μMORE was successfully applied to determine critical biocatalytic process parameters in a parallelized manner for the carboligation of benzaldehyde and acetaldehyde to (S)-2-hydroxy-1-phenylpropan-1-one with less than 5μg of benzoylformate decarboxylase from Pseudomonas putida immobilized on magnetic beads. Here, one run of the device in six parallelized glass reactors took only 2-3h for an immobilized enzyme with very low activity (∼2U/mg). The optimized parameter set was finally tested in a 10mL enzyme membrane reactor, demonstrating that the μMORE provides a solid data base for biocatalytic process optimization.
IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany. Electronic address: jussendaniel@aol.com.
Daniel
D
Jussen
ZEA-1: Engineering and Technology, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany. Electronic address: h.soltner@fz-juelich.de.
Helmut
H
Soltner
IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany. Electronic address: b.stute@fz-juelich.de.
Birgit
B
Stute
IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany. Electronic address: w.wiechert@fz-juelich.de.
Wolfgang
W
Wiechert
IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany. Electronic address: e.von.lieres@fz-juelich.de.
Eric
E
von Lieres
IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany. Electronic address: ma.pohl@fz-juelich.de.
Martina
M
Pohl
D019149:Bioreactors; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D004867:Equipment Design; D006863:Hydrogen-Ion Concentration; D059346:Magnets; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D008962:Models, Theoretical; D013696:Temperature
D004800:Enzymes, Immobilized
2-Hydroxy-1-phenylpropan-1-one (PubchemCID 101121); Acetaldehyde (PubchemCID 177); Benzaldehyde (PubchemCID 240); Biocatalytic process; CC bond formation; Immobilized enzyme; Magnetic beads; Microfluidic reactor; Process parameter optimization