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Applied and Environmental Microbiology, April 2007, p. 2079-2084, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.02826-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

L-Valine Production with Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum{triangledown}

Bastian Blombach,1 Mark E. Schreiner,1,{dagger} Jirí Holátko,1,{ddagger} Tobias Bartek,2 Marco Oldiges,2 and Bernhard J. Eikmanns1*

Institute of Microbiology and Biotechnology, University of Ulm, D-89069 Ulm,1 Institute of Biotechnology 2, Research Center Jülich, D-52425 Jülich, Germany2

Received 5 December 2006/ Accepted 26 January 2007

Corynebacterium glutamicum was engineered for the production of L-valine from glucose by deletion of the aceE gene encoding the E1p enzyme of the pyruvate dehydrogenase complex and additional overexpression of the ilvBNCE genes encoding the L-valine biosynthetic enzymes acetohydroxyacid synthase, isomeroreductase, and transaminase B. In the absence of cellular growth, C. glutamicum {Delta}aceE showed a relatively high intracellular concentration of pyruvate (25.9 mM) and produced significant amounts of pyruvate, L-alanine, and L-valine from glucose as the sole carbon source. Lactate or acetate was not formed. Plasmid-bound overexpression of ilvBNCE in C. glutamicum {Delta}aceE resulted in an approximately 10-fold-lower intracellular pyruvate concentration (2.3 mM) and a shift of the extracellular product pattern from pyruvate and L-alanine towards L-valine. In fed-batch fermentations at high cell densities and an excess of glucose, C. glutamicum {Delta}aceE(pJC4ilvBNCE) produced up to 210 mM L-valine with a volumetric productivity of 10.0 mM h–1 (1.17 g l–1 h–1) and a maximum yield of about 0.6 mol per mol (0.4 g per g) of glucose.

* Corresponding author. Mailing address: Institute of Microbiology and Biotechnology, University of Ulm, 89069 Ulm, Germany. Phone: 49 (0)731 50 22707. Fax: 49 (0)731 50 22719. E-mail: bernhard.eikmanns{at}uni-ulm.de.

{triangledown} Published ahead of print on 9 February 2007.

{dagger} Present address: R&D Women's Health, Europe, Johnson & Johnson GmbH, D-42289 Wuppertal, Germany.

{ddagger} Present address: Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídenská 1083, CZ-14220 Praha 4, Czech Republic.

Applied and Environmental Microbiology, April 2007, p. 2079-2084, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.02826-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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