Abstract
We recently engineered the wild type of Corynebacterium glutamicum for the growth-decoupled production of L-valine from glucose by inactivation of the pyruvate dehydrogenase complex and additional overexpression of the ilvBNCE genes, encoding the L-valine biosynthetic enzymes acetohydroxyacid synthase, isomeroreductase, and transaminase B. Based on the first generation of pyruvate-dehydrogenase-complex-deficient C. glutamicum strains, a second generation of high-yield L-valine producers was constructed by successive deletion of the genes encoding pyruvate:quinone oxidoreductase, phosphoglucose isomerase, and pyruvate carboxylase and overexpression of ilvBNCE. In fed-batch fermentations at high cell densities, the newly constructed strains produced up to 410 mM (48 g/l) L-valine, showed a maximum yield of 0.75 to 0.86 mol/mol (0.49 to 0.56 g/g) of glucose in the production phase and, in contrast to the first generation strains, excreted neither pyruvate nor any other by-product tested.
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Acknowledgement
We thank Lothar Eggeling for providing plasmids pJC4ilvBNC, pJC4ilvBNCD, and pJC4ilvBNCE and Brigitte Bathe (Evonik Degussa) for providing plasmid pK18mobsacB pgidel. We are grateful to Andreas Karau and Robert Gerstmeir (Degussa AG) for valuable discussions. We thank Konstanze Fleischer for technical assistance. The support of the Fachagentur Nachwachsende Rohstoffe of the BMVEL (grant 04NR004/22000404) is gratefully acknowledged.
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Blombach, B., Schreiner, M.E., Bartek, T. et al. Corynebacterium glutamicum tailored for high-yield L-valine production. Appl Microbiol Biotechnol 79, 471–479 (2008). https://doi.org/10.1007/s00253-008-1444-z
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DOI: https://doi.org/10.1007/s00253-008-1444-z