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Updated: Jun 7, 2025

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
From molasses to purified α-ketoglutarate with engineered Corynebacterium glutamicum
Lars Halle1, Daniela Höppner2, Marvin Doser2
1Institute of Bio- and Geosciences - IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52425 Jülich, Germany; Institute of Biotechnology, RWTH Aachen University, Worringer Weg 3, 52074 Aachen, Germany; Bioeconomy Science Center (BioSC), Forschungszentrum Jülich GmbH, 52425 Jülich, Germany.
Abstract:
α-ketoglutarate (AKG) is a valuable dicarboxylic acid with multiple applications in the food, pharmaceutical, and chemical industries. Its chemical synthesis is associated with toxic by-products, low specificity, and high energy input. To create a more environmentally friendly and sustainable alternative, a microbial production process for AKG was developed. Four potential producer strains were generated by metabolic engineering of Corynebacteriumglutamicum and characterized on defined glucose/sucrose media as well as molasses, a side stream from sugar beet processing. While strain C.glutamicum PO6-iolT1Δgdh ΔgltB mscCG' ΔodhA was not able to grow on defined media it outperformed all predecessor variants on molasses. Successful scale-up into a fed-batch bioreactor process with molasses yielded 96.2g AKG with a conversion yield of 0.64gg-1. Finally, downstream processing by liquid-liquid extraction with ethyl acetate enabled product purification with an extraction efficiency of 87 % and an AKG purity of > 93 %.

