Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 79, Issue 3, Pages 471-479Publisher
SPRINGER
DOI: 10.1007/s00253-008-1444-z
Keywords
Corynebacterium glutamicum; L-valine production; pyruvate dehydrogenase complex; pyruvate : quinone oxidoreductase; phosphoglucose isomerase; pyruvate carboxylase
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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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