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Authors: Hermann, Maria
Teleki, Attila
Weitz, Sandra
Niess, Alexander
Freund, Andreas
Bengelsdorf, Frank Robert
Dürre, Peter
Takors, Ralf
Title: Identifying and engineering bottlenecks of autotrophic isobutanol formation in recombinant C. ljungdahlii by systemic analysis
Issue Date: 2021 Zeitschriftenartikel 14 Frontiers in bioengineering and biotechnology 9 (2021), No. 647853
ISSN: 2296-4185
Abstract: Clostridium ljungdahlii (C. ljungdahlii, CLJU) is natively endowed producing acetic acid, 2,3-butandiol, and ethanol consuming gas mixtures of CO2, CO, and H2 (syngas). Here, we present the syngas-based isobutanol formation using C. ljungdahlii harboring the recombinant amplification of the “Ehrlich” pathway that converts intracellular KIV to isobutanol. Autotrophic isobutanol production was studied analyzing two different strains in 3-L gassed and stirred bioreactors. Physiological characterization was thoroughly applied together with metabolic profiling and flux balance analysis. Thereof, KIV and pyruvate supply were identified as key “bottlenecking” precursors limiting preliminary isobutanol formation in CLJU[KAIA] to 0.02 g L-1. Additional blocking of valine synthesis in CLJU[KAIA]:ilvE increased isobutanol production by factor 6.5 finally reaching 0.13 g L-1. Future metabolic engineering should focus on debottlenecking NADPH availability, whereas NADH supply is already equilibrated in the current generation of strains.
Appears in Collections:04 Fakultät Energie-, Verfahrens- und Biotechnik

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