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Applied and Environmental Microbiology, May 2002, p. 2246-2250, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2246-2250.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Linking Central Metabolism with Increased Pathway Flux: L-Valine Accumulation by Corynebacterium glutamicum

Eva Radmacher,1 Adela Vaitsikova,2 Udo Burger,1,{dagger} Karin Krumbach,1 Hermann Sahm,1 and Lothar Eggeling1*

Institut für Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany,1 BIOTIKA, 976 13 Slovenská Lupa, Slovakia2

Received 19 November 2001/ Accepted 12 February 2002

Mutants of Corynebacterium glutamicum were made and enzymatically characterized to clone ilvD and ilvE, which encode dihydroxy acid dehydratase and transaminase B, respectively. These genes of the branched-chain amino acid synthesis were overexpressed together with ilvBN (which encodes acetohydroxy acid synthase) and ilvC (which encodes isomeroreductase) in the wild type, which does not excrete L-valine, to result in an accumulation of this amino acid to a concentration of 42 mM. Since L-valine originates from two pyruvate molecules, this illustrates the comparatively easy accessibility of the central metabolite pyruvate. The same genes, ilvBNCD, overexpressed in an ilvA deletion mutant which is unable to synthesize L-isoleucine increased the concentration of this amino acid to 58 mM. A further dramatic increase was obtained when panBC was deleted, making the resulting mutant auxotrophic for D-pantothenate. When the resulting strain, C. glutamicum 13032{Delta}ilvA{Delta}panBC with ilvBNCD overexpressed, was grown under limiting conditions it accumulated 91 mM L-valine. This is attributed to a reduced coenzyme A availability and therefore reduced flux of pyruvate via pyruvate dehydrogenase enabling its increased drain-off via the L-valine biosynthesis pathway.

* Corresponding author. Mailing address: Institut für Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany. Phone: 49 2461 61 5132. Fax: 49 2461 61 2710. E-mail: l.eggeling{at}fz-juelich.de.

{dagger} Present address: Institut für Biochemie, Universität zu Köln, D-57674 Cologne, Germany.

Applied and Environmental Microbiology, May 2002, p. 2246-2250, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2246-2250.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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