Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum

Mahr R, Gaetgens C, Gaetgens J, Polen T, Kalinowski J, Frunzke J (2015)
Metabolic Engineering 32: 184-194.

Journal Article | Published | English

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Abstract
Adaptive laboratory evolution has proven a valuable strategy for metabolic engineering. Here, we established an experimental evolution approach for improving microbial metabolite production by imposing an artificial selective pressure on the fluorescent output of a biosensor using fluorescence-activated cell sorting. Cells showing the highest fluorescent output were iteratively isolated and (re-) cultivated. The L-valine producer Corynebacterium glutamicum Delta aceE was equipped with an L-valine-responsive sensor based on the transcriptional regulator Lrp of C. glutamicum. Evolved strains featured a significantly higher growth rate, increased L-valine titers (similar to 25%) and a 3-4-fold reduction of by-product formation. Genome sequencing resulted in the identification of a loss-of-function mutation (UreD-E188*) in the gene ureD (urease accessory protein), which was shown to increase L-valine production by up to 100%. Furthermore, decreased L-alanine formation was attributed to a mutation in the global regulator GlxR. These results emphasize biosensor-driven evolution as a straightforward approach to improve growth and productivity of microbial production strains. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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Mahr R, Gaetgens C, Gaetgens J, Polen T, Kalinowski J, Frunzke J. Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum. Metabolic Engineering. 2015;32:184-194.
Mahr, R., Gaetgens, C., Gaetgens, J., Polen, T., Kalinowski, J., & Frunzke, J. (2015). Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum. Metabolic Engineering, 32, 184-194.
Mahr, R., Gaetgens, C., Gaetgens, J., Polen, T., Kalinowski, J., and Frunzke, J. (2015). Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum. Metabolic Engineering 32, 184-194.
Mahr, R., et al., 2015. Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum. Metabolic Engineering, 32, p 184-194.
R. Mahr, et al., “Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum”, Metabolic Engineering, vol. 32, 2015, pp. 184-194.
Mahr, R., Gaetgens, C., Gaetgens, J., Polen, T., Kalinowski, J., Frunzke, J.: Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum. Metabolic Engineering. 32, 184-194 (2015).
Mahr, Regina, Gaetgens, Cornelia, Gaetgens, Jochem, Polen, Tino, Kalinowski, Jörn, and Frunzke, Julia. “Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum”. Metabolic Engineering 32 (2015): 184-194.
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