High Cell Density Cultivation Combined with high Specific Enzyme Activity: Cultivation Protocol for the Production of an Amine Transaminase from Bacillus megaterium in E. coli
ChemBioChem, vol. 25, pp. e202400006
Abstract
High cell density cultivation is an established method for the production of various industrially important products such as recombinant proteins. However, these protocols are not always suitable for biocatalytic processes as the focus often lies on biomass production rather than high specific activities of the enzyme inside the cells. In contrast, a range of shake flask protocols are well known with high specific activities but rather low cell densities. To overcome this gap, we established a tailor-made fed-batch protocol combining both aspects: high cell density and high specific activities of heterologously produced enzyme. Using the example of an industrially relevant amine transaminase from Bacillus megaterium, we describe a strategy to optimize the cultivation yield based on the feed rate, IPTG concentration, and post-induction temperature. By adjusting these key parameters, we were able to increase the specific activity by 2.6-fold and the wet cell weight by even 17-fold compared to shake flasks. Finally, we were able to verify our established protocol by transferring it to another experimenter. With that, our optimization strategy can serve as a template for the production of high titers of heterologously produced, active enzymes and might enable the availability of these catalysts for upscaling biocatalytic processes.
Authors 7
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Forschungszentrum Jülich · RWTH Aachen University
Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
RWTH Aachen University Aachen Biology and Biotechnology (ABBt) Worringer Weg 1 52074 Aachen Germany
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Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
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Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
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Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
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DSM (Netherlands) · Forschungszentrum Jülich · DSM-Firmenich AG (Switzerland)
Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
dsm-firmenich Alexander Fleminglaan 1 2613 AX Delft The Netherlands
DSM-Firmenich AG, Lab Automation & Screening, NETHERLANDS
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Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
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Dörte Rother Aachen
Forschungszentrum Jülich · RWTH Aachen University
Affiliation as printed
Forschungszentrum Jülich GmbH Institute of Bio- and Geo Science 1 (IBG-1): Biotechnology Wilhelm-Johnen-Straße 52425 Jülich Germany
RWTH Aachen University Aachen Biology and Biotechnology (ABBt) Worringer Weg 1 52074 Aachen Germany
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